WO2015191716A2 - Procédés, dispositifs et systèmes pour l'analyse d'échantillons - Google Patents

Procédés, dispositifs et systèmes pour l'analyse d'échantillons Download PDF

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Publication number
WO2015191716A2
WO2015191716A2 PCT/US2015/035112 US2015035112W WO2015191716A2 WO 2015191716 A2 WO2015191716 A2 WO 2015191716A2 US 2015035112 W US2015035112 W US 2015035112W WO 2015191716 A2 WO2015191716 A2 WO 2015191716A2
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WIPO (PCT)
Prior art keywords
assay
sample
assays
initial
subsequent
Prior art date
Application number
PCT/US2015/035112
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English (en)
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WO2015191716A3 (fr
Inventor
Daniel Young
Elizabeth Holmes
Original Assignee
Theranos, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Theranos, Inc. filed Critical Theranos, Inc.
Priority to CA2951558A priority Critical patent/CA2951558A1/fr
Priority to MX2016016234A priority patent/MX2016016234A/es
Priority to EP15807539.0A priority patent/EP3155420A4/fr
Publication of WO2015191716A2 publication Critical patent/WO2015191716A2/fr
Publication of WO2015191716A3 publication Critical patent/WO2015191716A3/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/06—Investigating concentration of particle suspensions
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/1023—Microstructural devices for non-optical measurement
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1429—Signal processing
    • G01N15/1433—Signal processing using image recognition
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/06—Investigating concentration of particle suspensions
    • G01N15/075—Investigating concentration of particle suspensions by optical means
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/06—Investigating concentration of particle suspensions
    • G01N2015/0687—Investigating concentration of particle suspensions in solutions, e.g. non volatile residue
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1006—Investigating individual particles for cytology
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N2015/1488—Methods for deciding
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N2015/1493—Particle size
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N2015/1497—Particle shape

Definitions

  • Methods, devices, and systems for the analysis of biological samples are provided which may be configured to perform multiple assays on a sample or samples, comprising an initial assay or multiple initial assays, and further comprising a subsequent assay or subsequent assays, the performance of, and/or order of performance of, such subsequent assay or assays being contingent on the results of such initial assay(s).
  • a subsequent assay or such subsequent assays may be performed in a contingent manner, wherein the performance of a subsequent assay depends on the results of one or more prior assay.
  • whether or not a subsequent assay is performed at all may be contingent on the results of a. prior assay.
  • the order of performance of subsequent assays, or of steps in a subsequent assay may be contingent on the results of a prior assay.
  • the choice of a subsequent assay, from among a plurality of possible subsequent assays may be contingent on the results of a prior assay.
  • the method of performing a subsequent assay may be contingent on the results of a prior assay.
  • the timing of the performance of a subsequent assay may be contingent on the results of a prior assay.
  • the choice of a reagent used in a subsequent assay may be contingent on the results of a prior assay.
  • the choice of a method of detection used in a subsequent assay may be contingent on the results of a prior assay.
  • the choice of whether or not to use a second biological sample in a subsequent assay may be contingent on the results of a prior assay in which a first biological sample was assayed.
  • the choice of whether or not to obtain a second biological sample may be contingent on the results of a prior assay of a first biological sample. In embodiments, the choice of whether or not to obtain a second biological sample for use in a subsequent assay may be contingent on the results of a prior assay of a first biological sample.
  • analysis of a biological sample may comprise assaying the sample, or a portion of the sample, for the presence of an analyte.
  • An analyte is the subject of an analy sis.
  • An analyte may be a natural constituent of a biological sample, or may be an element, compound, material, or cell not normally found in a biological sample that may be the subject of analysis.
  • An analyte may comprise a chemical compound, e.g., a small molecule, a protein, a nucleic acid, or other compound, present in the sample.
  • An analyte may comprise a physical or chemical characteristic of a sample, or of a portion of the sample.
  • An analyte may comprise a marker or physical or chemical characteristic of a cell or virus in a sample, or in a portion of the sample.
  • An analyte may comprise a cell or virus, or portion thereof, in a sample, or in a portion of the sample.
  • a method of testing a biological sample comprises:
  • an initial assay for the presence of an analyte in said biological sample, whereby an initial result is obtained, wherein said initial assay may provide a negative result indicating that the presence of said analyte is not detected in the biological sample, or may- provide a positive result indicating that the presence of the analyte is detected in the biological sample;
  • a method of testing a biological sample comprises:
  • an initial assay in a device for an analyte in said biological sample, whereby an initial result is obtained, wherein said initial assay may provide a negative result indicating that the presence of said analyte is not detected, or is detected at a normal level, in the biological sample, or may provide a positive result indicating that the presence of the analyte is detected, or is detected at an abnormal level, in the biological sample;
  • such subsequent assay may be a type of assay selected from the group consisting of antibody-based assays, nucleic acid assays, general chemistry assays, and cytometric assays, m embodiments, the subsequent assay may comprise an assay of a different type than the initial assay type. In embodiments, such subsequent assays may comprise assays that are more sensitive for (he detection of said analyte than said initial assay. In embodiments, the analyte to be detected by such subsequent assays may comprise a different analyte than the analyte to be detected by said initial assay.
  • such an initial assay may comprise the use of a detector to obtain said initial result wherein said detector is selected from an optical detector, a pH detector, an electrochemical detector, a temperature sensor, an ion-sensitive electrode, a radiation detector, and other detectors.
  • a subsequent assay may comprise the use of a detector to obtain a further result, wherein said detector is selected from an optical detector, a pH detector, an electrochemical detector, a temperature sensor, an ion- sensitive electrode, a radiation detector, and other detectors.
  • Protocols disclosed herein comprise protocols for performance of a sequence of assays
  • protocols may include contingent assays, whose performance is dependent on the results of prior assays, wherein a result of a first assay, if it meets a criterion, triggers, alters, or prevents the performance of a subsequent assay.
  • a subsequent assay is automatically performed if the results of an initial assay, or other prior assays, meet the criterion or criteria.
  • such a subsequent assay may not be performed in the absence of prior results which meet the criterion.
  • the performance of a subsequent assay may be contingent on two, or three, or more criteria.
  • Protocols include contingent sequences of assays, where the performance of a subsequent assay is determined by the outcome of an assay performed prior to the performance of the subsequent assay .
  • a protocol including one or more contingent sequences of assay s may be requested at the time the assays are ordered.
  • a protocol including one or more contingent sequences of assays may be determined at the time the assays are ordered.
  • a protocol including one or more contingent sequences of assays may be requested at the time the assays are ordered, and the particulars of the sequence, and of the contingent steps, may be determined at a time after the time the assays are ordered.
  • a protocol including one or more assays, whose performance or sequence of performance is contingent on the result of an initial assay may include instructions, or a protocol, for the acquisition of a biological sample.
  • a protocol may direct or require thai a biological sample of sufficient volume or amount be acquired in order to provide sufficient biological sample for the performance of a contingent assay, if the contingent assay is needed.
  • such a protocol may direct or require that a biological sample be divided into aliquots for the performance of a contingent assay, if the contingent assay is needed.
  • such a protocol may direct or require that a biological sample be diluted, in order to provide sufficient biological sample for the performance of a contingent assay , if the contingent assay is needed. In embodiments, such a protocol may direct or require that a biological sample, or portion or dilution thereof, be retained for use in the performance of a contingent assay, if the contingent assay is needed.
  • an assay may utilize a diluted sample (e.g., a diluted biological sample).
  • a diluted sample e.g., a diluted biological sample.
  • an assay may include instructions, or a protocol, for the dilution of a biological sample.
  • a protocol including one or more assays, whose performance or sequence of performance is contingent on the result of an initial assay may include instructions, or a protocol, for the dilution of a biological sample.
  • such dilution may provide a diluted sample diluted by at least about 10-fofd as compared to the original sample; or may provide a diluted sample diluted by at least about 30-fold as compared to the original sample; or may provide a diluted sample diluted by at least about 50-fold as compared to the original sample; or may provide a diluted sample diluted by at least about 100-fold as compared to the original sample; or may provide a diluted sample diluted by at least about 1000-fofd, or more, as compared to the original sample.
  • such a protocol may direct or require that a biological sample, or portion thereof, including diluted portions thereof, be retained for the performance of a contingent assay, if the contingent assay is needed,
  • a contingent assay is automatically run if the contingent assay is indicated on the test order or other instruction, regardless of the results from any initial assay.
  • Such contingent assay(s) may be run concurrently with or even before results are available from any initial assay(s).
  • alternative protocols may result in additional usage of reagent(s), diluent(s), and/ or other materials, the workflow of parallel, concurrent, or continuous processing may be beneficial in some situations, such as but not limited excess system availability while processing the initial assays or waiting for initial assays to complete.
  • an initial assay may comprise a less sensitive assay, and a subsequent assay may comprise a more sensitive assay; in embodiments, performance of a subsequent assay may be contingent on the results of the initial assay.
  • an initial assay may comprise a nucleic acid assay performed under a first condition
  • a subsequent assay may comprise a nucleic acid assay performed under a second condition, where the second condition comprises more stringent nucleic acid assay conditions than the first condition (e.g., the second condition comprises a higher temperature than the first condition, or the second condition comprises a lower ionic strength than the first condition, or the second condition comprises a denaturing agent (such as formamide) not present, or present at a lower concentration, in the first condition).
  • an initial assay that comprises a nucleic acid assay may be performed under moderately stringent conditions
  • a subsequent assay may comprise a nucleic acid assay performed under high stringent conditions.
  • an initial assay may comprise a first type of assay, and a subsequent assay, contingent on the results of the initial assay may comprise a second type of assay; for example, an initial assay may comprise an antibody-based assay, and a subsequent assay may comprise a nucleic acid assay .
  • Applicant further discloses methods of testing a biological sample in a device, comprising: performing an initial assay in said device for an analyte in said biological sample, whereby an initial result is obtained, wherein said initial assay may provide a negative result indicating that the presence of said analyte is not detected, or is detected at a normal level, in the biological sample, or may provide a positive result indicating that the presence of the analyte is detected, or is detected at an abnormal level, in the biological sample; performing further testing of said biological sample, wherein a subsequent assay is performed in said device on said biological sample regardless of the results of said initial assay; and reporting the results of said subsequent assay of said biological sample contingent upon the results of said initial assay.
  • the methods further comprising reporting the results of said initial assay.
  • the results of said subsequent assay are not reported if said initial assay provides a negative result, and wherein the results of said subsequent assay are reported if said initial assay provides a positive result.
  • the results of said subsequent assay are not reported if said initial assay provides a positive result, and wherein the results of said subsequent assay are reported if said initial assay provides a negative result.
  • the subsequent assay is an assay for the same ana!yte as said initial assay, and the subsequent assay is a more sensitive assay than said initial assay.
  • the biological sample upon which the subsequent assay is performed is obtained before the results of said initial assay are obtained.
  • the initial assay and the subsequent assay are performed o portions of the same biological sample.
  • at least one of said portions of said biological sample is a diluted portion of the biological sample.
  • the subsequent assay comprises an assay of a type selected from the group of assay types consisting of antibody-based assays, nucleic acid assays, general chemistry assays, and cytometric assays.
  • the subsequent assay comprises a different type of assay than the initial assay .
  • the analyte to be detected by said subsequent assay comprises a different analyte than the analyte detected by said mitial assay.
  • the initial assay comprises measurement of an analyte
  • said subsequent assay comprises a cytometric assay.
  • the methods, compositions, devices, and systems provide rapid tests, which require only small biological samples, and thus provide advantages over other methods, compositions, assays, devices, and systems.
  • Devices and systems disclosed herein are configured to perform such rapid assays which require only small amounts of sample, such as only small amounts of sample, urine, sputum, tears, material obtained from a nasal swab, throat swab, cheek swab, or other biological sample. Accordingly, the methods, devices, and systems provide rapid tests, which require only small biological samples, and thus provide advantages over other methods, compositions, assays, devices, and systems.
  • a biological sample may comprise a sample selected from blood, serum, plasma, a throat sw r ab, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid, tears, stool, mucus, sweat, earwax, oil, a glandular secretion, semen, vaginal fluid, interstitial fluids derived from tumorous tissue, ocular fluids, breath, hair, finger nails, skin, biopsy tissue, placental fluid, amniotic fluid, cord blood, lymphatic fluids, cavity fluids, sputum, pus, microbiota, meconium, breast milk, and other secretions or excretions.
  • such an initial assay and such a subsequent assay may be performed on different biological samples.
  • the biological sample upon which the subsequent assay is performed may be obtained before the results of said initial assay are obtained, in embodiments, the biological sample upon which the subsequent assay may be performed is obtained after the results of said initial assay are obtained.
  • Applicant discloses herein methods in which a subsequent assay- is performed within a short amount of time from the time at which the biological sample tested by the subsequent assay was accepted within the device.
  • the initial assay and the subsequent assay are each performed on at least a portion of the same biological sample; in such embodiments, the subsequent assay is performed within a short amount of time from the time at which an initial biological sample was accepted within the device.
  • a subsequent assay is performed if an initial assay is performed.
  • a subsequent assay is performed on a biological sample in the same device within a short amount of time from the time of accepting a sample within said device, wherein said short amount of time is a time prior to the performance of the subsequent assay.
  • the same sample is used for the performance of both the initial assay and the subsequent assay, or portions of the same sample are used for the performance of both the initial assay and the subsequent assay.
  • Such a short amount of time may be, for exampl e, a short amount of time consisting of about 3 hours or less; about 2 hours or less; about 1 hour or less; about 50 minutes or less; about 45 minutes or less; about 40 minutes or less; about 35 minutes or less; about 30 minutes or less; about 25 minutes or less; about 20 minutes or less; about 15 minutes or less; about 10 minutes or less; about 5 minutes or less; about 4 minutes or less; about 3 minutes or less; about 2 minutes or less; and about 1 minute or less.
  • the biological sample upon which the subsequent assay is performed is obtained after the results of said initial assay are obtained; in such embodiments, the subsequent assay is performed within a short amount of time from the time at which a further biological sample was accepted within the device.
  • a further biological sample may be obtained following the time at which an initial biological sample is obtained.
  • a further biological sample is obtained from a subject during a single session in which biological samples are obtained.
  • a further biological sample is obtained from a subject during a second, or subsequent, session following the session during which the initial biological sample was obtained.
  • a second, or subsequent, session may follow the session during which the initial biological sample was obtained by a time period of about 10 minutes or less; or about 20 minutes or less; or about 30 minutes or less; or about 40 minutes or less; or about 50 minutes or less; or about one hour or less; or about two hours or less; or about three hours or less; or other time period.
  • the assays and methods disclosed herein may be performed on a device, or on a system, for processing a sample.
  • the assays and methods disclosed herein can be readily incorporated into and used in an automated assay device, and in an automated assay system.
  • a device as disclosed herein may be suitable for the detection, identification, or measurement of an analyte or characteristic in a biological sample.
  • a device as disclosed herein may be an automated assay device, suitable for automated detection, identification, or measurement of an analyte or characteristic in a biological sample.
  • Such automated detection, identification, or measurement of an analyte or characteristic in a biological sample may include, for example, detection, identification, or measurement of an analyte or characteristic in a biological sample according to a protocol performed by the automated assay device: a processor may be used in the performance of assays according to such a protocol.
  • the automated detection, identification, or measurement of a characteristic in a biological sample may include the automated detection, identification, or measurement of a characteristic of a cell in the biological sample, such as may be detected, identified, or measured, e.g., by a cytometric assay, and may include the automated detection, identification, or measurement of a morphological characteristic of a cell in the biological sample.
  • a device for assaying a sample for the presence of analyte in a sample may comprise: a fluid handling system for transporting at least a portion of a biological sample; and a detector effective to detect or measure an analyte or characteristic.
  • a detector may comprise one or more of an optical detector, a pH sensor, an electrochemical detector, a radiation detector, a temperature sensor, and other sensors.
  • An optical detector may comprise one or more of a camera, a photomultiplier, a photodiode, a spectrophotometer, and other optical elements.
  • a detector may comprise an imaging device or imaging system, and may include optical elements such as lenses, filters, mirrors, gratings, polarizers, and other optical elements.
  • a device for assaying a sample for the presence of analyte in a sample may comprise a system for transporting at least a portion of a sample.
  • a device for assaying a sample for the presence of analyte in a sample may comprise a sy stem for transporting a reagent.
  • a system for transporting at least a portion of a sample, or for transporting a reagent may comprise a fluid handling system.
  • a fluid handling system of a device for assaying a sample for the presence of analyte in a sample may be configured to transport at feast a portion of a biological sample and also be configured to transport a reagent.
  • a device for assaying a sample for the presence of analyte in a sample may be configured to dilute at least a portion of a sample with a reagent.
  • a device for assaying a sample for the presence of analyte in a sample may be configured to mix a reagent with at least a portion of a sample.
  • a device comprising a fluid handling system for transporting at least a portion of a biological sample may comprise a means for contacting a biological sample; or may comprise a means for accepting a biological sample; or may comprise means for storing a biological sample.
  • a biological sample may be provided by a cartridge.
  • reagents for assays to be performed on a biological sample may be provided by a cartridge.
  • all reagents required for the assays to be performed on a biological sample may be provided by a cartridge.
  • consumables including pipette tips, mixing vessels, cuvettes, and other containers, implements, and materials, may be provided by a cartridge; in embodiments, all such materials required for the assays to be performed on a biological sample may be provided by a cartridge.
  • a cartridge may hold a container in which a biological sample is contained.
  • a container configured to hold a biological sample may be configured to store a biological sample, and may be configured to allow access to said biological sample by a fluid handling system.
  • Access by a fluid handling system to a biological sample may be effective to allow transport of at least a portion of the biological sample; to allow mixing of the biological sample; to allow addition of a reagent to at least a portion of the biological sample; or to allow division of said biological sample into two or more portions
  • a cartridge may hold a biological sample, and reagents for use in the assays to be performed on the sample; in embodiments, a biological sample and all reagents required for the assays to be performed on a biological sample may be provided by a single cartridge.
  • a cartridge may hold a biological sample, reagents, and consumables, including pipette tips, mixing vessels, cuvettes, and other containers, implements, and materials for use in the assays to be performed on the sample; in embodiments, a single cartridge may carry a biological sample, and all reagents and all consumables required for the assays to be performed on a biological sample.
  • sy stems are provided which include devices configured to detect the presence of an analyte in a biological sample.
  • systems are provided which include devices configured to detect the presence of an analyte in a biological sample, and a cartridge containing reagents for assays.
  • systems are provided which include devices configured to detect the presence of an anaiyte in a biological sample, and a cartridge containing reagents for assays, and consumable items for assays.
  • a device or system may further comprise a communication assembly, which may comprise a display element and/or a communication element effective to report the results of said detection and/or measurement.
  • a communication assembly such as a display element and/or communication element, may be suitable for two-way communication.
  • a communications assembly may be configured to communicate data obtained from assaying a sample
  • a device for assaying a sample for the presence of anaiyte in a sample may comprise other elements and assemblies, including, without limitation, a heating assembly, a cooling assembly, a sonicator, and other elements and assemblies,
  • systems as disclosed herein may include a communication assembly for transmitting or receiving a protocol based on the anaiyte to be detected or based on other analytes to be detected by the device or system.
  • an assay protocol may be changed based on results previously obtained from a sample from a subject, or based on results previously obtained from a different sample from the subject.
  • a communication assembly may comprise a channel for communicating information from said device to a computer, said wherein said channel is selected from a computer network, a telephone network, a metal communication link, an optical communication link, and a wireless communication link.
  • systems as disclosed herein may transmit signals to a central location, or to an end user, and may include a communication assembly for transmitting such signals.
  • Systems as disclosed herein may be configured for updating a protocol as needed or on a regular basis.
  • Applicant discloses devices configured to measure an anaiyte in a biological sample according to a method disclosed herein.
  • Devices configured to measure analytes in a biological sample according to a method disclosed herein may be configured to measure analytes from a biological sample that comprises no more than about 1000 uL of sample, or no more than about 500 ⁇ , of sample, no more than about 250 ⁇ , of sample, or no more than about 200 ⁇ , of sample, or no more than about 150 ⁇ , of sample, or no more than about 100 ⁇ , of sample, or no more than about 75 ⁇ .
  • ⁇ _ of sample refers to the volume, in ⁇ ,, of a sample
  • a biological sample that comprises no more than about 100 ⁇ . of sample has a volume of no more than about 100 ⁇ ...
  • Such devices e.g., devices configured to measure an anaiyte in a biological sample according to a method disclosed herein
  • Devices disclosed herein may be configured to perform an assay for the measurement of a first anaiyte and also to perform an assay for the measurement of a second anaiyte in the biological sample.
  • performance of an assay for the measurement of a second anaiyte in the biological sample may be contingent on the results of an assay for the measurement of a first anaiyte in the biological sample.
  • devices disclosed herein may be configured to perform an assay for the measurement of an anaiyte and also to perform an assay comprising the measurement of a morphological characteristic of a blood cell in the blood sample.
  • Devices disclosed herein may be configured to perform an assay for the measurement of a first anaiyte and also to perform an assay comprising the measurement of another blood anaiyte, e.g., a v itamin, a hormone, a drug or metabolite of a drag, or other anaiyte.
  • Such devices may be configured wherein the assays, or the order of performance of assays, that are performed by said device may be altered by communication with another device.
  • Applicant also discloses systems comprising a device as disclosed herein.
  • the system comprises a device that is configured to perform an assay for the measurement of a first anaiyte and also to perform an assay for the measurement of another anaiyte in the blood sample.
  • the system comprises a device that is configured to perform an assay for the measurement of an anaiyte and also to perform an assay for the measurement of a morphological characteristic of a blood cell in the blood sample.
  • assays, or the order of performance of assays, that are performed by said device may be altered by communication with another device.
  • Fig. 1 shows representative images of blood cells from a sample of whole blood, illustrating various types of images that may be obtained using different imaging techniques and dyes.
  • Fig. 2 shows a representative composite image of several cell-types in whole blood, including images of a monocyte, a lymphocyte, an eosinophil, and a neutrophil.
  • Fig. 3A shows a plots of monocytes identified and quantified by the cytometric assays described herein.
  • Fig. 3B shows a plots of basophils identified and quantified by the cytometric assays described herein.
  • Fig. 3C shows a plots of lymphocytes identified and quantified by the cytometric assays described herein.
  • Fig. 3D shows a plots of neutrophils and eosinophils identified and quantified by the cytometric assays described herein.
  • Fig. 4A shows a plot demonstrating that cytometric methods as disclosed herein identify different cell types consistent with such identification by other methods.
  • Fig. 4A shows plots of numbers of white blood ceils ("WBCs") from blood samples.
  • WBCs white blood ceils
  • Fig. 4B shows a plot demonstrating that cytometric methods as disclosed herein identify different cell types consistent with such identification by other methods.
  • Fig. 4B shows plots of numbers of red blood cells ("RBCs) from blood samples.
  • Fig. 4C shows a plot demonstrating that cytometric methods as disclosed herein identify different cell types consistent with such identification by other methods.
  • Fig. 4C shows plots of numbers of platelets from blood samples.
  • Fig. 4D shows a plot demonstrating that cytometric methods as disclosed herein identify different cell types consistent with such identification by other methods.
  • Fig. 4D shows plots of numbers of neutrophils from blood samples.
  • Fig. 4E shows a plot demonstrating that cytometric methods as disclosed herein identify different cell types consistent with such identification by other methods.
  • Fig. 4E shows plots of numbers of monocytes from blood samples.
  • Fig. 4F shows a plot demonstrating that cytometric methods as disclosed herein identify different cell types consistent with such identification by other methods.
  • Fig. 4F shows plots of numbers of ly mphocytes from blood samples.
  • Fig, 5A shows a first side of an order sheet including Istings of test panels and of reflex tests according to the methods disclosed herein.
  • Fig. 5B shows a second side of an order sheet including listings of test panels and reflex tests according to the methods disclosed herein.
  • Fig. 6A shows a further example of an order sheet listing individual tests, test panels, test groupings, and including lists of reflex tests according to the methods disclosed herein.
  • Fig. 6B shows shows a listing of panel components and capped price offerings for the list shown in Fig, 6A.
  • Fig. 7A shows a further example of an order form listing individual tests, test panels, test groupings, and including lists of reflex tests according to methods disclosed herein,
  • Fig. 7B shows a listing of panel components and capped price offerings for the list shown in Fig. 7A. CPT codes are listed for the panels and panel components.
  • Fig, 7C shows a further listing of panel components and capped price offerings for the list shown in Fig, 7A. CPT codes are listed for the panels and panel components.
  • Fig. 8A shows a further example of an order form listing individual tests, test panel s, test groupings, and including lists of reflex tests according to methods disclosed herein.
  • Fig. 8B shows a listing of panel components and capped price offerings for the list shown in Fig. 8 A. CPT codes are listed for the panels and panel components.
  • automated assay device and “automated assay system” refer to devices and systems which, once started in operation, are capable of running substantially without hitman control or intervention, and may be run according to one or more protocois which govern the particulars of ihe operation of the device or sy stem
  • automated assay devices and systems are typically configured to facilitate the collection of a sample; to prepare a sample for an assay (e.g., a clinical test); to effect a chemical reaction between a sample and one or more reagents; to produce, detect, or measure a chemical or physical characteristic or process in a sample: to observe, image, detect, identify, or measure a chemical or physical characteristic in a sample; and, in general, to perform assays and obtain assay results from a sample (e.g., a biological sample).
  • an assay e.g., a clinical test
  • reagents e.g., a chemical reaction between a sample and one or more reagents
  • An automated assay device may be configured to process a sample; to obtain data from a sample; to transmit daia obtained from a sample; to analyze data from a sample; and to perform other operations on and with a sample (e.g., a biological sample).
  • An automated assay device may be configured to communicate with another device, or a laboratory, or an individual affiliated with a laboratory, to analyze data obtained from a sample (e.g., a biological sample).
  • an automated assay device or system may be equivalently termed a sample processing device or system, a sample analysis device or system, an automatic assay device or system, an automatic sample processing device or system, an automatic sample analysis device or system, or other such terms.
  • the terms ''consumable” and “consumables” refer to implements and materials useful in the performance of assays, and include pipette tips, mixing vessels, cuvettes, and other containers, implements, and materials which may be used during an assay for the detection or measurement of an analyte or characteristic in or of a biological sample.
  • An automated sample analysis device or system may use consumables during the performance of an assay. Consumables may be carried on a cartridge, and may be provided for use by an automated sample analysis device, e.g., for use in the performance of an assay.
  • the term "assay” and its grammatical equivalents refers to tests, measurements, observations, and other experimental procedures which may be applied to a sample for detection of an analyte, identification of an analyte, and measurement of the amounts of an analyte in a sample.
  • Assays may be physical assays which detect, identify, or measure a physical property of a sample; assays may be chemical assays, which detect, identify, or measure a chemical property of a sample, or perform chemical reactions in or with a sample; and include assays which use optical, electrical or electronic, chemical, or other means of detection and measurement.
  • test and “tests” and grammatical equivalents thereof are used interchangeably with the term “assay”, and refer to measurements, observations, and other experimental procedures which may be applied to a sample for detection of an analyte, identification of an analyte, and measurement of the amounts of an analyte in a sample.
  • the terms "panel”, “assay panel”, and grammatical equivalents refers to two or more assays that may be ordered, and that may be performed, together as a group, on a sample or samples for detection of an analyte, identification of an analyte, and measurement of the amounts of an analyte in a sample, or for the detection of two or more analyses, or identification of two or more analytes, or measurements of the amounts of two or more analytes in a sample or samples.
  • the assays grouped together in a panel are typically, although not necessarily, directed to markers or characteristics indicative of the same disease or disorder (e.g., to markers or characteristics indicative of celiac disease), or are directed to markers or characteristics indicative of the same condition (e.g., to markers or characteristics indicative of pregnancy, or of cardiovascular disease, or of liver disorders), or are directed to markers or characteristics indicative of similar diseases or disorders (e.g., to markers or characteristics indicative of a respiratory disease or a digestive system disease or disorder), or are directed to markers or characteristics indicative of diseases or disorders having similar symptoms (e.g., to markers or characteristics indicative of a viral infection, a bacterial infection, a blood disease or disorder, or a thyroid disease or disorder), or are directed to markers or characteristics indicative of diseases or disorders having similar causes or risk factors (e.g., sexually transmitted diseases), or are directed to markers or characteristics that may occur together in a subject (e.g., to markers or characteristics indicative of a plurality of infectious diseases).
  • markers or characteristics indicative of the same disease or disorder e.
  • an "analyte” is the subject of an assay or analysis, the presence or amounts of which are to be determined by the performance of the assay.
  • An analyte may be a natural constituent of a biological sample; an analyte may be free in a fluid sample or solution, or may be bound to another compound (e.g., to a carrier protein), may be present on a cell, or may be present in a cell; an analyte may be a compound not normally found in a biological sample, such as a drug; a metabolite of a drug or other compound; an infectious agent (e.g., a virus, bacteria, or other foreign organism or material); a toxin; or other compound, element, cell, or cellular marker that may be the subject of analysis.
  • An analyte may comprise a cell, or virus, or portion thereof; and may comprise a physical or chemical characteristic of a cell, vims, tissue, or portion thereof.
  • An assay for an analyte is a test or procedure directed at detecting the presence of that analyte, or determining the amount of that analyte, or identifying that analyte, or characterizing that analyte, or otherwise obtaining information about that analyte, in a sample.
  • characteristics refers to a dimension, property, marker, or other attribute of a sample or component of a sample (e.g., a biological sample); in embodiments, characteristics may include the presence or distribution of markers, such as identifying markers, on a cell; characteristics may include morphological characteristics of a cell, whether before treatment or after treatment of that cell; characteristics may include physical characteristics (e.g., size, volume, shape, viscosity, mechanical properties, or other physical characteristics) including optical characteristics (e.g., color, reflectance, absorbance, polarization, light scattering, or other optical characteristics; characteristics may include chemical characteristics, including, e.g., presence or concentration of elements or compounds, pH, chemical reactivity, and other characteristics; and other characteristics. Such characteristics may be detected, identified, measured, or otherwise assayed on a sample as a whole, or on components of a sample (e.g., cells within a biological sample).
  • An assay for a characteristic is a test or procedure directed at detecting the presence of that characteristic, or identifying that characteristic, or quantifying that characteristic, or characterizing that characteristic, or otherwise obtaining information about that characteristic, in a sample (e.g., a biological sample).
  • An assay for an analyte or characteristic is a test or procedure directed at detecting the presence of that analyte or characteristic, or determining the amount of that analyte or that characteristic, or identifying that analyte or characteristic, or characterizing that analyte or characteristic, or otherwise obtaining information about that analyte or characteristic, in a sample (such as a biological sample).
  • negative result refers to a result of an assay in which the presence of the target analyte is not detected, or is detected at a normal level, in the biological sample.
  • the term "positive result” and grammatical equivalents thereof refers to a result of an assay in which the presence of the target analyte is detected, or is detected at an abnormal level, in the biological sample.
  • reflex test and "reflex assay” refer to an assay (equivalently, a test) the performance of which is contingent on a result of a previous test (which may be termed an "initial test” or an “initial assay”).
  • a reflex assay may be termed a subsequent assay (e.g., when performed following an initial assay). Whether or not the reflex test is performed may be determined by a result of an initial test.
  • the timing of the performance of a reflex test may be determined by a result of an initial test.
  • the order of performance of a reflex test, or reflex tests, may be determined by a result of an initial test
  • a "subsequent test” may be a reflex test. For example, where a subsequent test is performed only upon obtaining a particular result, or one of a particular group of results, then that subsequent test may be a reflex test. For example, where a subsequent test is reported to a subject (or to a clmican, doctor, or other health care professional regarding that subject), only upon obtaining a particular result, or one of a particular group of results, then that subsequent tes t may be a reflex test.
  • the term "contingent” refers to dependence on a prior event, result, condition, or state.
  • a criterion e.g., the occurrence of an act; the outcome of a test; the existence of a state or condition; or the satisfaction of a criterion of any kind
  • the performance of that act is conditional upon the criterion, and that act will occur or be performed, or not, or will occur or be performed in a particular way, depending upon that criterion.
  • one fonn of reflex test is one in which a positive result from a general test triggers an automatic test of a more specific nature; for example, an influenza reflex test which is positive for the presence of influenza virus may automatically trigger a more specific test which will identify the particular strain of influenza present in a sample from the subject, or will determine whether or not a particular strain of influenza is present in a sample from the subject.
  • the term "contingent assay” refers to an assay the performance of which (or failure to perform) depends on a prior event or condition.
  • a contingent assay may be one that is performed only if the result of a prior assay satisfies a criterion (or criteria); a contingent assay may be one that is not performed if the result of a prior assay satisfies a criterion (or criteria); a contingent assay may be one that is performed in one way if the result of a. prior assay satisfies a criterion (or criteria), but is performed in a different way if the criterion (or criteria) is (are) not met.
  • a singl e contingent assay may depend upon a single criterion; or, a single contingent assay may depend upon multiple criteria. Multiple contingent assays may depend upon a single criterion; or, multiple contingent assays may depend upon multiple criteria,
  • a step in an assay may be a "contingent step", i.e., one that is contingent upon the occurrence or outcome of a prior assay , or a prior step of an assay (including the assay in which a contingent step occurs).
  • CPT refers to "current procedural terminology” and the term “C-PT code” refers to the code used to identify a diagnostic, treatment, or other clinical procedure.
  • Current Procedural Terminology codes (“CPT codes”) are used to describe clinical procedures, such as medical, surgical, and diagnostic procedures. Thus, CPT codes identify a procedure, including procedures used to determine a diagnosis, while other codes may be used to identify diagnoses.
  • CPT codes are often required in order to obtain reimbursement for a procedure by insurance companies and other third-party payers. For example, CPT codes may be required in order to obtain reimbursement from a government agency (e.g., in order to obtain Medicare or Medicaid reimbursement).
  • ICD International Classification of Diseases
  • ICD code refers to the code used to identify a particular disease or condition.
  • International Classification of Diseases codes (“ICD codes") are revised periodically; current revisions of ICD codes which are still typically used include ICD-9 and ICD-10.
  • ICD codes typically identify a disease, and not a particular procedure associated with such a disease.
  • the ICD codes have also been modified to include procedures as well as diagnoses; thus, ICD-9-CM and ICD-10-CM include diagnosis codes, and ICD-9-PCS and ICD- 10-PCS include procedure codes.
  • ICD codes are used to describe a disease or disorder suffered by a subject, and may be used or required in order to obtain reimbursement for a procedure by insurance companies and other third-parry payers.
  • ICD-9-CM codes may be required in order to obtain reimbursement from a government agency (e.g., in order to obtain Medicare or Medicaid reimbursement).
  • abbreviations and acronyms used herein, including in the figures, are those in common usage in the art.
  • the abbreviations and acronyms as used herein typically identify biomarkers, biochemical or physiological quantities or measurements, assays for those biomarkers, or specific assays or assay types.
  • Such abbreviations and acronyms include “Ab” which stands for antibody; “Ag” which stands for antigen; “ABO/RhD” stands for a blood- typing test to determine type A, B, or O and Rhesus factor positive or Rhesus factor negative (Rh+ or Rh-); “ALP” is an acronym for alkaline phosphatase; “ALT” is an acronym for alanine aminotransferase; “ASP” is an acronym for aspartase aminotransferase; “BUN” is an acronym for blood urea nitrogen; “CBC” is an acronym for complete blood count; “CONF” refers to a “confirmatory test”, i.e., a subsequent test used to confirm a possible finding from an initial test; each confirmatory test is specific for the initial test which it is designed to confirm (for example, "HIV W/'CONF” indicates that a positive HIV test reflexes to a confirmatory differential test between HIV-1 and HIV -2); “CRP” is an acronym for C-
  • MHC histocompatibility class II antigens
  • HsCRP high sensitivity C-reactive protein assay
  • HSV herpes simplex virus (including, e.g., forms 1 and 2, HSV1 and HSV2)
  • LDL low-density lipoprotein
  • LH low-density lipoprotein
  • PSA prostate specific antigen
  • PT prothrombin time
  • PT71NR prothrombin time/international normalized ratio
  • PTT is an acronym for partial thromboplastin time
  • Qual stands for qualitative; “Quant” stands for quantitative;
  • RPC is an acronym for red blood cell;
  • RNA is an acronym for ribonucleic acid;
  • RPR stands for rapid plasma reagin;
  • Sed Rate stands for sedimentation rate;
  • T3 stands for trifluoride transfer protein
  • MHC histocompatibility class II antigens
  • reflex to culture indicates that, where the results of a test indicate or require a subsequent test (a "reflex" test, e.g., for confirmation, for quantification, or for turiher specification of the target detected, or for other purpose), that reflex testing includes a culture assay, e.g., an assay in which a portion of a sample is placed in a culture medium or culture dish and maintained under culture conditions for further testing.
  • a culture assay e.g., an assay in which a portion of a sample is placed in a culture medium or culture dish and maintained under culture conditions for further testing.
  • Reflex to culture & susceptibility indicates that, where the results of a test indicate or require a subsequent test (a "reflex" test, e.g., for confirmation, for quantification, or for further specification of the target detected, or for other purpose) that reflex testing includes a culture assay and a susceptibility assay (i.e., an assay in which different anti-microbiai agents are applied to culture assays to determine which anti-microbial agents would be most effective in combatting an infection identified by the culture assay).
  • a susceptibility assay i.e., an assay in which different anti-microbiai agents are applied to culture assays to determine which anti-microbial agents would be most effective in combatting an infection identified by the culture assay.
  • Microalbumiii/Creatinine Urine Random indicates that the (urine) sample may be taken at any time during the day or night.
  • the term “Complete” in the phrase “Urinalysis, Complete” indicates that microscopy is used in the analysis of a urine sample in addition to the other automated aspects of the urinalysis (e.g., chemical and optical examination).
  • antibody and “'antibodies” refer to complete antibodies and to functional (i.e., specifically binding) fragments and variants thereof. Antibody may be abbreviated “Ab”. An antibody specifically binds to a target antigen; an antigen may be abbreviated "Ag” (e.g., "HBsAg” indicates hepatitis B surface antigen).
  • Antibodies may be identified by the target molecule to which they specifically bind; for example, an "anti-HiV” antibody binds HIV virus particles with high specificity, and an "anti-CD41” antibody binds CD41 antigens with high specificity.
  • IgA refers to antibodies of the immunoglobulin A type
  • IgD refers to antibodies of the immunoglobulin D type
  • IgE refers to antibodies of the immunoglobulin E type
  • IgG refers to antibodies of the immunoglobulin G type
  • IgM refers to antibodies of the immunoglobulin M type.
  • antibody-based assay refers to an assay for an analyte which may be found in a biological sample, using specific binding of the analyte by an antibody, antibody fragment, or antibody mimetic (e.g., immunoadhesin, aptamer, or other construct which specifically binds to a target with little or no cross-reactivity with other compounds) to detect the presence of, and, if desired, quantify the amounts of, an analyte in a sample.
  • an antibody, antibody fragment, or antibody mimetic e.g., immunoadhesin, aptamer, or other construct which specifically binds to a target with little or no cross-reactivity with other compounds
  • antibody-based assay includes immunoassays (including ELISAs), rece tor-based assays, and other assays which utilize specific binding between a receptor and a ligand to detect, identify, or quantify an analyte.
  • a Western blot may be termed an antibody-based assay.
  • An ap tamer is a nucleic acid molecule capable of binding to a target molecule.
  • the nucleic acid may be a deoxyribonucleic acid, a ribonucleic acid, a linked peptide nucleic acid, or other nucleic acid, analog, or derivative thereof.
  • the generation and use of aptamers is known in the art; see, e.g., U.S. Patent No. 5,475,096. Aptamers may be used with, or in place of, antibodies in assays in which require binders which specifically bind to target molecules in a sample (e.g., "antibody-based assays").
  • immunoadhesin designates an antibody-like molecule which combines the binding specificity of a heterologous protein (an “adhesin”) with the effector functions of immunoglobulin constant domains.
  • the immunoadhesins comprise a fusion of an amino acid sequence with the desired binding specificity which is other than the antigen recognition and binding site of an antibody (i.e., is "heterologous"), and an immunoglobulin constant domain sequence.
  • the adhesin part of an immunoadhesin molecule typically is a contiguous amino acid sequence comprising at least the binding site of a receptor or a ligand.
  • the immunoglobulin constant domain sequence in the immunoadhesin may be obtained from any immunoglobulin, such as IgG-1, lgG-2, IgG-3, or IgG4 subtypes, TgA. (including IgA-1 and TgA -2), IgE, gD or TgM, Immunoadhesms reported in the literature include fusions of the T cell receptor (Gascoigne et al, Proc. Natl. Acad. Sci.
  • Immunoadhesms may be used with, or in place of, antibodies in assays in which require binders which specifically bind to target molecules in a sample (e.g., "antibody-based assays”) .
  • cytometric assay refers to an assay which detects, or identifies, or quantities cells or particles in a sample, typically by optical means (e.g., by microscopy). For example, cells or particles may be counted in a sample, or a field of view within a sample, to provide a number, or density, or other numerical value regarding cells or particles in a sample. Size, or optical intensity, or other characteristic of cells or particles in a sample may be measured or characterized. Ceils in a sample assayed in a cytometric assay- may be labeled, or otherwise treated, to enhance their identification or to ease the differentiation between cells and cell types.
  • cell surface markers may be labeled and optically identified by exposure of cells to antibodies or antibody fragments directed at particular cell surface antigens, where the antibodies or antibody fragments are labeled with fluorescent dyes or other identifiable markers.
  • aptamers, immunoadhesms, nucleic acid molecules, nucleic acid mimetics, dyes, and other probes or labels specific for cellular features, organelles, molecules, or characteristics may also be used to identify and characterize cells in a cytometric assay.
  • ''measurement of a morphological characteristic of a cell or particle refers to a cytometric assay directed at identifying, quantifying, or characterizing the size, shape, or other physical characteristic of a cell, particle, or group of cells or particles in a sample. Observation, measurement, or
  • characterization of the appearance of a ceil or particle is one type of measurement of a morphological characteristic.
  • Measurement or characterization of the size of a cell or particle may refer to the measurement or characterization of the diameter, cross-sectional area, volume, or other characteristic of a cell or particle.
  • Measui'ement or characterization of the shape of a cell or particle may refer to the measurement or characterization of the symmetry (or asymmetry), or presence or number of protrusions, smoothness (or roughness) of a cellular surface, or other characteristic of a cell or particle.
  • Measurement or characterization of brightness, or other optical property, across a dimension of a cell or particle comprises measurement of a morphological characteristic of a cell or particle. Other morphological characteristics may also be observed, measured and characterized.
  • label or “marker” or the phrases “detectable label” and “marker moiety” when used herein refer to a detectable compound or composition which is conjugated directly or indirectly to the antibody so as to generate a "labeled” antibody.
  • the label may be deiectable by itself (e.g.
  • radioisotope labels or fluorescent labels or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition which is detectable (a label may be, e.g., a dye, a fluorescent moiety, a luminescent moiety, a chemiluminescent moiety, an enzymatic label, a magnetic label, a paramagnetic label, a contrast agent, a nanoparticle, a radioisotope, an epitope tag, biotin, streptavidin, or a quencher).
  • a label may be, e.g., a dye, a fluorescent moiety, a luminescent moiety, a chemiluminescent moiety, an enzymatic label, a magnetic label, a paramagnetic label, a contrast agent, a nanoparticle, a radioisotope, an epitope tag, biotin, streptavidin, or a quencher).
  • Dyes include, for example, fluorescent dyes that intercalculate with double-stranded DNA include, for example, SYBR GoldTM, SYBR Green ITM, SYBR Green HTM, ethidium bromide, BlueViewTM, methylene blue, DAP1, and acridine orange.
  • Further fluorescent dyes include, for example, CA.L Fluor Gold, CAL Fluor Orange, Quasar 570, CAL Fluor Red 590, CAL Fluor Red 610, CAL Fluor Red 610, CAL Fluor Red 635, Quasar 670 (Biosearch Technologies), VIC, NED (Life Technologies), Cy3, Cy5, Cy5.5 (GE Healthcare Life Sciences), Oyster 556, Oyster 645 (Integrated DNA Technologies), LC red 610, LC red 610, LC red 640, LC red 670, LC red 705 (Roche Applies Science), Texas red, FAM, TET, HEX, JOE, TMR, and ROX.
  • Quenchers that may be used include, for example, DDQ-I, DDQ-1I (Eurogentec), Eclipse (Epoch Biosciences), Iowa Black FQ, Iowa Black RQ (Integrated DNA Technologies), BHQ-1, BHQ-2, BHQ-3 (Biosearch Technologies), QSY- 7, QSY-21 (Molecular Probes), and Dabcyl.
  • small molecule refers to a compound, typically a non-polymeric organic compound, that is smaller than a typical protein.
  • small molecules include acetylsalicylie acid (aspirin), caffeine, cholesterol, vitamin D, and other molecules.
  • a small molecule typically has a molecular weight below about 500 Daltons.
  • small molecules may be small organic molecules, and may be small inorganic molecules.
  • general chemistry assay refers to an assay for an element or compound which may be found in a biological sample, using chemical or physical reactions to detect the presence of, and, if desired, quantify the amounts of, an anaiyte in a sample.
  • a "general chemistry assay” may be directed at detecting a small molecule anaiyte (including ions and elements such as sodium or potassium), or directed at a chemical characteristic of a sample (e.g., H, O2 saturation, or other sample characteristic determined by chemical means).
  • nucleic acid assay refers to an assay for an nucleic acid (whether deoxyribonucleic acid (DNA) or ribonucleic acid (RNA)) which may be found in a biological sample; such assays typically use hybridization between a probe and the target nucleic acid to detect the presence of, and, if desired, qitanttfy the amounts of, an analvte in a sample.
  • DNA deoxyribonucleic acid
  • RNA ribonucleic acid
  • Nucleic acid assays include, for example, Northern blots, Southern blots, polymerase chain reaction (PGR) assays of all types, including, e.g., PGR, reverse transcriptase PGR (RT- PCT), quantitative PGR (qPCR) (see, e.g. U.S. Patent No. 4,683,202;), and other assays.
  • Nucleic acid assays may utilize thermal cycling; nucleic acid assays include isothermal assays which do not require thermal cycling, or include steps which do not require thermal cycling (such as, e.g., loop-mediated isothermal amplification ("LAMP”) as described, e.g., in U.S. Patent No.
  • LAMP loop-mediated isothermal amplification
  • Nucleic acid assays may be performed at one or more levels of stringency.
  • nucleic acid assays may be performed at high stringency (e.g., under "stringent conditions” or “high stringency conditions”).
  • nucleic acid assays including nucleic acid assays discussed herein, may be performed at moderate stringency (e.g., under "moderately stringent conditions”).
  • nucleic acid assays, including nucleic acid assays discussed herein may be performed at low stringency (e.g., under "low stringent conditions”).
  • Hybridization generally depends on the ability of denatured DNA or RNA to reanneal when complementary strands are present in an environment below their melting temperature.
  • stringency of hybridization reactions see Ausubel et al., Current Protocols in Molecular Biology, Wiley Interscience Publishers, (1995).
  • Stringent conditions or “high stringency conditions”, as defined herein, may be identified by those that: (1 ) employ low ionic strength and high temperature for washing, for example 0.015 M sodium chioride/0.0015 M sodium citrate/0.1% sodium dodecyl sulfate at 50°C; (2.) employ during hybridization a denaturing agent, such as formamide, for example, 50% (v/v) formamide with 0.1 % bovine serum albumm/0.1. % Ficoll/0.1% polyvinylpyrrolidone/50 mM sodium phosphate buffer at pH 6.5 with 750 mM sodium chloride.
  • a denaturing agent such as formamide, for example, 50% (v/v) formamide with 0.1 % bovine serum albumm/0.1.
  • washing solution and hybridization conditions e.g., temperature, ionic strength and % SDS
  • washing solution and hybridization conditions e.g., temperature, ionic strength and % SDS
  • An example of moderately stringent conditions is overnight incubation at 37°C. in a solution comprising: 20% Formamide, 5 x SSC (150 mM NaCl, 15 mM trisodium citrate). 50 mM sodium phosphate (pH 7.6). 5 x Denhardt's solution.
  • Low stringent conditions may include the use of washing solution and hybridization conditions (e.g., temperature, ionic strength and % SDS) even less stringent than those for moderately stringent conditions; such conditions include lower temperatures (temperatures lower than 37 °C, such as, e.g., 32 °C, or 30 °C, or lower) and higher salt concentrations than are typically used for moderately stringent conditions.
  • washing solution and hybridization conditions e.g., temperature, ionic strength and % SDS
  • blood and “whole blood” refer to blood as it exists within an animal and as directly obtained from a subject in a blood sample. Blood contains red blood cells, white blood cells, proteins such as albumin, globulins, and clotting factors, salts, water, and other constituents.
  • plasma and "blood plasma” refer to the liquid portion of blood (e.g., a blood sample) that remains after the removal of blood cells. Red blood cells and white blood cells may be removed by centrifugation of a blood sample, leaving plasma above the pelleted cells in the bottom of the centrifuge tube. Plasma retains blood clotting factors, and is obtained from anti-eoagulated blood samples.
  • serum refers to the liquid portion of blood that remains after blood is allowed to clot, and the clot is removed. Serum differs from plasma in that serum lacks clotting factors: since clotting requires fibrin, thrombin, and other proteins, which form and remain part of a blood clot, serum lacks these proteins while plasma contains them.
  • a "finger-stick” refers to: i) the act of making a small puncture in the skin of a subject, allowing a small amount (e.g., a droplet, or one, two, or a few drops) of blood to flow and become available for collection; ii) ihe puncture itself; and iii) ihe blood collected thereby. Blood may be liberated in a finger-stick, for example, by use of a lancet or other sharp implement effective to pierce the skin of a subject.
  • the amount of blood may be about 250 ⁇ _, or less, or about 200 s iiL or less, or about 150 ⁇ iL or less, or about 100 ⁇ . or less, or about 50 ⁇ _ or less, or about 25 ⁇ iL or less, or about 15 ⁇ 1_ or less, or about 10 ⁇ or less, or about 10 ⁇ iL or less, or about 5 ⁇ or less, or about 3 ⁇ or less, or about i ⁇ or less).
  • Blood from a finger-stick may be collected, e.g., by needle, syringe, capillary tube, or other method. Blood from a finger-stick may be collected for transport to another location; for storage prior to use or analysis; for immediate use; or for a combination of the same.
  • biological sample refers to a fluid, tissue, or other material collected from a subject.
  • biological samples can include but are not limited to, blood, serum, plasma, a throat swab, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid, tears, stool, mucus, sweat, earwax, oil, a glandular secretion, semen, vaginal fluid, interstitial fluids derived from tumorous tissue, ocular fluids, breath, hair, finger nails, skin, biopsy tissue, placental fluid, amniotic fluid, cord blood, lymphatic fluids, cavity fluids, sputum, pus, microbiota, meconium, breast milk, and other secretions or excretions.
  • Biological samples may include nasopharyngeal wash, or other fluid obtained by washing a body cavity or surface of a subject, or by washing a swab following application of the swab to a body cavity or surface of a subject.
  • Nasal swabs, throat swabs, stool samples, hair, finger nail, ear wax, breath, and other solid, semi-solid, or gaseous samples may be processed in an extraction buffer, e.g., for a fixed or variable amount of time, prior to their analysis. The extraction buffer or an aliquot thereof may then be processed similarly to other fluid samples if desired.
  • tissue samples of the subject may include but are not limited to, connective tissue, muscle tissue, nervous tissue, epithelial tissue, cartilage, cancerous sample, or bone.
  • the sample may be obtained from a human or animal.
  • the sample may be obtained from a mammal, vertebrate, such as murines, simians, humans, farm animals, sport animals, or pets.
  • the sample may be obtained from a living or dead subject.
  • the sample may be obtained fresh from a subject or may have undergone some form of pre-processing, storage, or transport.
  • portion and portions include, without limitation, aliquots (which may be of equal volume, or may be of unequal volume); dilutions (in which a sample, or a portion thereof, is mixed with a diluent to form a dilution of the sample); fractions (e.g., a fraction of whole blood, such as serum, or plasma, or a pellet formed by centrifugation); and combinations thereof.
  • Methods, devices, and systems disclosed herein may be used to provide and perform a "reflex test” or “reflex assay” in which a second assay is, or subsequent assays are, performed contingent on the results of an initial test or tests.
  • "reflex” refers to the performance of a subsequent test pursuant to performing, or to obtaining the results of, an initial test.
  • a reflex algorithm may dictate the performance of a nucleic acid assay to detect the presence of, or to quantify the amount of, that disease- causing organism in a sample obtained from the subject.
  • the nucleic acid assay is termed a "reflex test” or “reflex assay” with respect to the antibody assay.
  • the antibody assay is said to "reflex” to the nucleic acid assay.
  • a reflex test may be performed upon obtaining negative results from an initial test.
  • a negative result may indicate that a subject does not suffer from a suspected disease or condition that is suggested by a particular symptom, and indicates that a different assay directed at a different suspected disease or condition should be performed.
  • a reflex test for the different suspected disease or condition may be performed, as a result of obtaining the negative results from the initial test.
  • a reflex test may be performed upon obtaining positive results from an initial test.
  • an initial test may be, e.g, an HIV test.
  • positive results of an antibody -based assay for HIV e.g., an HTV- 1 test, or an HIV- 2 test
  • positive results of an antibody -based assay for HIV on a sample from a subject may trigger the testing of a sample from the same subject using a nucleic acid assay for HIV (e.g., a nucleic acid assay for HIV-1 test, or a nucleic acid assay for H1V-2 test).
  • a positive result in an antibody-based HIV assay for HIV may trigger testing of a sample from the same subject in a nucleic acid assay.
  • a positive result in an initial nucleic acid assay for HIV may trigger testing of a sample from the same subject by a subsequent, higher stringency nucleic acid assay for HIV.
  • a positive result in an antibody -based HIV assay e.g., an ELISA or other assay
  • Western blot also an antibody-based assay
  • a positive result in an antibody-based HIV assay may trigger testing of a sample from the same subject in a cytometric assay.
  • a positive result in a nucleic acid assay for HIV may trigger testing of a sample from the same subject in a cytometric assay.
  • a positive result in an antibody- based HIV assay for HIV may trigger testing of a sample from the same subject in a general chemistry assay.
  • a positive result in a nucleic acid assay-based HIV assay for HIV may trigger testing of a sample from the same subject in a general chemistry assay.
  • a positive result in a cytometric assay indicative of symptoms of HIV may trigger testing of a sample from the same subject in an antibody-based assay, or a nucleic acid assay, or in a general chemistry assay, or a combination of one or more of these assays.
  • a positive result in any assay indicative of symptoms of HIV may trigger testing of a sample from the same subject in a cyiometric assay , an antibody-based assay, or a nucleic acid assay , or in a general chemistry assay, or a combination of one or more of these assays.
  • an initial assay may comprise a complete blood cell count test, comprising testing a blood sample for white blood cells, where a white blood ceil count outside the normal range (e.g., below about 2000 cells per microliter (p.L) for males older than 12 years of age is outside the normal range) may trigger a reflex test comprising further testing of the blood of the subject.
  • a white blood ceil count outside the normal range e.g., below about 2000 cells per microliter (p.L) for males older than 12 years of age is outside the normal range
  • a reflex test comprising further testing of the blood of the subject.
  • further testing of the blood may be an automatic test performed by a cytometer on a device as disclosed herein.
  • a reflex cytometric test may comprise identifying and/or quantifying white blood cells in the sample by ceil type, using a cytometer.
  • Such a cytometer test may be an automatic cytometer test, and may not require the intervention of any person to perform the cytometric assay.
  • further testing of the blood may be a "Manual" test performed by a trained person.
  • a reflex test comprising further testing of the blood may comprise both an automatic test performed by a cytometer on a device and a "Manual" test performed by a framed person.
  • a positive test for hepatitis may trigger the further testing of a sampie from the same subject.
  • an initial test for hepatitis may comprise a test for analytes indicative of compromised liver function, e.g., increased serum aminotransferase levels, or increased alkaline phosphatase levels, or increased gamma- giutamyl transpeptidase levels, or increased bilirubin, or other blood marker which may be altered by hepatitis.
  • such further testing of a sampie from the subject may comprise an antibody-based assay.
  • such further testing of a sample from the subject may comprise a nucleic acid assay.
  • such further testing of a sample from the subject may comprise an automatic test performed by a cytometer on a device as disclosed herein.
  • a positive test for hepatitis C may trigger the further testing of a sampie from the same subject.
  • an initial test for hepatitis C may comprise a test for the presence of antibodies to hepatitis C virus.
  • such further testing of a sample from the subject may be a nucleic acid assay for the presence of nucleic acids indicative of hepatitis C virus.
  • an initial test for hepatitis C may comprise an antibody-based assay for the presence of epitopes (antibody targets) indicative of the presence of hepatitis C virus in the sampie.
  • such further testing of a sampie from the subject may be a nucleic acid assay for the presence of nucleic acids indicative of hepatitis C virus.
  • an initial test for hepatitis C may comprise a test for the presence of nucleic acids indicative of hepatitis C virus.
  • such further testing of a sample from the subject may be a nucieic acid assay for the presence of antibodies to hepatitis C vims.
  • a positive test for the presence of syphilis may trigger the further testing of a sample from the same subject.
  • an initial test for the presence of syphilis may comprise a nucleic acid test for the presence of nucleic acids indicative of syphilis bacteria.
  • a subsequent test for the presence of syphilis may comprise a higher stringency nucleic acid test for the presence of syphilis bacteria (as compared to the stringency of the initial nucleic acid test for syphilis).
  • a subsequent test for the presence of syphilis may comprise a Western blot test for the presence of syphilis bacteria.
  • an initial test for the presence of syphilis may comprise a test for the presence of antibodies to syphilis bacteria.
  • further testing of a sample from the subject may be an automatic test performed by a cytometer on a device as disclosed herein.
  • an automatic test performed by a cytometer on a device as disclosed herein may be a cytometric test using darkfield illumination of a blood sample from the patient, in order to determine and/or quantify the presence of syphilis bacteria.
  • such further testing may be a "Manual" test performed by a trained person.
  • An initial assay may comprise an assay for the presence of hepatitis B.
  • a sample obtained from a pregnant female may be tested for hepatitis B surface antigen (an antibody-based assay). If this initial test is found to be positive for the presence of hepatitis B surface antigen, a nucleic acid reflex test may be performed for a more sensitive and specific test for the hepatitis B vims; the reflex test may also provide quantification of the viral load in the sample.
  • the nucleic acid reflex test may be performed on a different sample than the antibody-based hepatitis B surface antigen assay; this different sample may be a portion of the original sample, obtained by dividing the original sample into portions, and retained for future use if needed; this different sample may be a separate sample, obta ined at the same, or nearly the same time, as the original sample, and retained for future use if needed; or this different sample may be a sample obtained after obtaining the original sample (e.g., after determination of the result of the ini tial test ).
  • a sample obtained from a pregnant female may be tested for hepatitis B surface antigen (an antibody-based assay). If this initial test is found to be positive for the presence of hepatitis B surface antigen, a reflex assay for the presence of syphilis may be performed.
  • a reflex syphilis assay may be, for example, a nucleic acid refl ex test for the presence of nucleic acids indicative of the presence of syphilis bacteria in the sample.
  • such a reflex syphilis assay may be an antibody -based reflex test for the presence of proteins indicative of the presence of syphilis bacteria in the sample.
  • such a reflex syphilis assay may be, for example, a cytometric assay for the presence of syphilis bacteria in the sample.
  • the reflex test may be performed on a different sample than the antibody-based hepatitis B surface antigen assay; this different sample may ⁇ be a portion of the original sample, obtained by dividing the original sample into portions, and retained for future use if needed; this different sample may be a separate sample, obtained at the same, or nearly the same time, as the original sample, and retained for future use if needed; or this different sample may be a sample obtained after obtaining the original sample (e.g., after determining the result of the initial test).
  • a urine sample may be assayed for the presence of blood, for the presence of nitrite, for the presence of more than trace amounts of protein, or for the presence of white blood cells (e.g., leukocytes) in the urine, in embodiments, an initial assay performed on a urine sample may be an antibody-based assay. If the results of such an antibody-based assay are found to be positive, then a reflex test may be performed, in which a sample of urine is subjected to cytometric testing. Such cytometric testing may be automated, or may be manual (i.e., involving observation of the sample by a technician using a microscope, or a camera, or both), or a combination of these.
  • cytometric testing may be automated, or may be manual (i.e., involving observation of the sample by a technician using a microscope, or a camera, or both), or a combination of these.
  • the cytometric reflex test may be performed on a different sample than the antibody-based initial assay; this different sample may be a portion of the original sample, obtained by dividing the original sample into portions, and retained for future use if needed; this different sample may be a separate sample, obtained at the same, or nearly the same time, as the original sample, and retained for future use if needed: or this different sample may be a sample obtained after obtaining the original sample (e.g., after determining the result of the initial test).
  • a urine sample may be assayed for the presence of blood, for the presence of nitrite, for the presence of more than trace amounts of protein, or for the presence of white blood cells (e.g., leukocytes) in the urine, as discussed above.
  • an initial assay performed on a urine sample may be an antibody-based assay. If the results of such an antibody -based assay are found to be positive, then a reflex test may be performed, in which an assay for the presence of bacteria is performed, using a sample of urine.
  • such an assay for the presence of bacteria in the urine comprises a bacterial culture, in which urine is applied to a substrate suitable for the culture of bacteria and the substrate incubated under conditions conducive to bacterial growth.
  • a bacterial culture reflex test may be performed on a different sample than the antibody-based initial assay; this different sample may be a portion of the original sample, obtained by dividing the original sample into portions, and retained for future use if needed; this different sample may ⁇ be a separate sample, obtained at the same, or nearly the same time, as the original sample, and retained for future use if needed; or this different sample may be a sample obtained after obtaining the original sample (e.g., after determining the result of the initial test).
  • all assays are performed automatically by a device or system, e.g., by a device or system as disclosed herein.
  • all assays are performed automatically by a device or system, e.g., by a device or system as disclosed herein.
  • all assays are performed automatically by a device or system, e.g., by a device or system as disclosed herein.
  • an initial assay is performed automatically by a device or system, e.g., by a device or system as disclosed herein, and a further sample is requested; upon loading such further sample in a device, at least one subsequent assay is performed automatically on the subsequent sample by a device or system, e.g., by a device or system as disclosed herein.
  • an initial assay is performed automatically by a device or system, e.g., by a device or system as disclosed herein, and a further sample is requested; upon loading such further sample in a device, all subsequent assays are performed automatically on the subsequent sample by a device or system, e.g., by a device or system as disclosed herein.
  • an initial assay is performed automatically by a device or system, e.g., by a device or system as disclosed herein, and a further sample is obtained automatically; at least one subsequent assay is performed on such subsequent sample automatically by a device or system, e.g., by a device or system as disclosed herein.
  • an initial assay is performed automatically by a device or system, e.g., by a device or system as disclosed herein, and a further sample is obtained automatically; all subsequent assays are performed on such subsequent sample automatically by a device or system, e.g., by a de vice or system as disclosed herein.
  • Reflex testing may be indicated, or may be suggested, or may be performed in response to an assay result.
  • the results of a first assay A may be determinative of whether or not a second assay B should be run; in this example, assay A is the initial assay, and assay B, contingent on the results of assay A, is the reflex assay.
  • Reflex testing may be indicated where the results of a first assay A determine that a second assay B should be run (e.g., in this case, the results of assay A indicate that assay B should be run).
  • Reflex testing may be suggested where the results of a first assay A determine that a second assay B should be run (e.g., in this case, where the results of assay A.
  • a display, a message, an alert, or a report, etc. may be provided (e.g., to the subject, to a health care professional, or other suitable party) suggesting that assay B should be run).
  • Reflex testing may be performed where the results of a first assay A determine that a second assay B should be run (e.g., in this case, the results of assay A indicate that assay B should be run).
  • the results of that assay may be provided (e.g., in a display , message, alert, or report) to the subject, a health care professional, or other suitable party.
  • reflex testing may be performed in response to an assay result.
  • the results of a first assay A may be determinative of whether or not a second assay B should be run; in this example, assay A. is the initial assay, and assay B, contingent on the results of assay A, is the reflex assay.
  • a cartridge may be pre-loaded with reagents required by assay A, and also preloaded with reagents necessary for assay B. If the result of assay A meets a predefined criterion initiating the reflex assay, then assay B is run with the same sample in the device.
  • the device protocol is planned to account for the possibility of running the reflex test (i.e., the necessary reagents are loaded into the cartridge, sufficient sample is obtained to perform both assay A and assay B, and sufficient sample is retained in reserve for running assay B if needed).
  • some protocol steps of assay B may be performed before the results for assay A are complete.
  • sample preparation can be completed in advance on the device.
  • a protocol including one or more assays, whose performance or sequence of performance is contingent on the result of an initial assay may include instructions, or a protocol, for the dilution of a biological sample.
  • such dilution may provide a diluted sample diluted by at least about 10-fold as compared to the original sample; or may provide a diluted sample diluted by at least about 20-fold as compared to the original sample; or may provide a diluted sample diluted by at least about 30-fold as compared to the original sample; or may provide a diluted sample diluted by at least about 50-fold as compared to the original sample; or may provide a diluted sample diluted by at least about 100- fold as compared to the original sample; or may provide a diluted sample diluted by at least about 200-fold as compared to the original sample; or may provide a diluted sample diluted by at least about 300-fold as compared to the original sample; or may provide a diluted sample diluted by at least about 500
  • a sample of sufficient size may be obtained from the subject sufficient to perform assay A and to perform assay B.
  • a sample may be obtained from the subject, and a portion of the sample may be diluted with a reagent (e.g., a diluent such as water, saline, a buffered solution, or other diluent) in order to provide sufficient volume of sample to perform one of assay A or assay B with undiluted sample, and to provide sufficeint diluted sample to perform one of assay A or assay B with diluted sample
  • a sample may be obtained from the subject, and all or a portion of the sample may be diluted with a reagent (e.g., a diluent such as water, saline, a buffered soluiion, or other diluent) in order to provide sufficient volume of diluted sample to perform assay A and to perform assay B.
  • a reagent e.g., a diluent such
  • a sample may be obtained from the subject, used to perform assay A, and stored in order to be available for the performance of assay B if indicated.
  • assay B is performed on the stored portion.
  • assay B is performed on a diluted sample, or on a diluted portion of a sample, where the stored portion is an undiluted portion, and the undiluted portion is diluted prior to performing assay B.
  • assay B is performed on a diluted sample, or on a diluted portion of a sample, where the stored portion is an undiluted sample, or undiluted sample portion.
  • a sample may be obtained from the subject, and divided into two portions, one of which may be used to perform assay A, and the other of which may be stored in order to be available for the performance of assay B if indicated. In embodiments, if the performance of assay B is indicated, assay B is performed on the stored portion. In embodiments, both portions of a sample di vided into two portions (one of which may be used to perform assay A, and the other of which may be used to perform assay B) may be treated equivalently. In embodiments, the portions of a sample divided into two portions (one of which may be used to perform assay A, and the other of which may be used to perform assay B) may be treated differently from each other.
  • a reflex assay may be performed with a second sample from the patient.
  • a second sample may be obtained from the subject following the satisfaction of the criterion or criteria by the results of assay A.
  • a result of assay A may trigger a request for a second, or a subsequent, sample.
  • a request for a second, or a subsequent, sample may comprise a message, e.g., a message displayed on the device, a message displayed on an interface, a message sent to an electronic address (e.g., an email, or an internet posting, or a tweet, or other address of an electronic messaging system), a message to the subject, or to an operator of a device, or to a health-care provider, or to a payer (e.g., an insurance company), or more than one of these messages, or to more than one of these individuals or entities.
  • a second or subsequent sample may comprise same type of sample as the initial sample.
  • a second or subsequent sample may comprise a different type of sample than the initial sample.
  • a second sample may be obtained from the subject prior to the satisfaction of the criterion or criteria by the results of assay A; for example, two samples may be obtained from a subject at the same time, or may be obtained at different times prior to obtaining the results from the initial assay A.
  • both samples may be treated equivalently.
  • both samples may be treated differently from each other,
  • Applicant discloses a method of testing a small-volume biological sample in a device, wherein said small-volume biological sample has a volume of less than about 250 microliters (fiL), the method comprising: placing said small-volume sample within the device; dividing said small-volume biological sample into at least a first and a second portion; retaining said second portion within the device for use in a subsequent assay; performing, in said device on said first portion or aliquot thereof, an init al assay for detecting or measuring an analyte or characteristic of the sample, whereby an initial result is obtained from the first portion, wherein: i) said initial result may comprise a negative result indicating that the presence of said analyte or characteristic is not detected or measured in the biological sample, or is detected or measured at a normal level in the biological sample, or ii) said initial result may comprise a positive result indicating that the presence of the analyte or characteristic is detected or measured in the biological sample, or is detected or measured at an abnormal
  • the performance of a subsequent assay is begun pursuant to the initial result within a short amount of time after placing the small- volume sample within the device, wherein said short amount of time is about two hours or less.
  • the performance of a subsequent assay is begun within a short time comprising about three hours, or about two hours, or about one hour, or about 45 minutes, or about 30 minutes, or about 20 minutes, or about 15 minutes, or about 10 minutes, or about 5 minutes, or about 4 minutes, or about 3 minutes, or about 2 minutes, or about 1 minute, or less.
  • the performance of a subsequent assay is completed within a short amount of time after placing the small-volume sample within the device, wherein said short amount of time is about two hours or less.
  • the performance of a subsequent assay is completed within a short time comprising about three hours, or about two hours, or about one hour, or about 45 minutes, or about 30 minutes, or about 20 minutes, or about 15 minutes, or about 10 minutes, or about 5 minutes, or about 4 minutes, or about 3 minutes, or about 2 minutes, or about 1 minute, or less.
  • the first portion of the small- volume sample is diluted prior to performance of the initial assay.
  • the second portion of the small-volume sample is diluted prior to performance of the subsequent assay.
  • both the first portion and the second portion of the small-volume sample are diluted prior to performance of the initial and the subsequent assays, respectively.
  • the first portion of the small-volume sample is diluted prior to performance of the initial assay.
  • the second portion of the small-v olume sample is diluted prior to performance of the subsequent assay.
  • both the fsrst portion and the second portion of the small-volume sample are diluted prior to performance of the initial and the subsequent assays, respectively.
  • a subsequent assay may comprise an assay for detecting or measuring the same analyte or characteristic as an initial assay, and such a subsequent assay may be more sensitive for the detection or measurement of the analyte or characteristic than the initial assay.
  • the analyte or characteristic to be detected or measured by a subsequent assay may comprise a different analyte or characteristic than the analyte or characteristic detec ted or measured by an initial assay.
  • an initial assay may comprise an assay of an assay type selected from the group of assay types consisting of antibody-based assays, nucleic acid assays, general chemistry assays, and cytometric assays
  • a subsequent assay may comprise a different type of assay than said initial assay.
  • an initial assay may comprise a nucleic acid assay
  • a subsequent assay may comprise an assay selected from an antibody-based assay, a general chemistry assay, and a cytometric assay.
  • an initial assay may comprise an antibody assay, and a subsequent assay may comprise an assay selected from a nucleic acid assay, a general chemistry assay, and a cytometric assay.
  • an initial assay may comprise a general chemistry assay, and a subsequent assay may comprise an assay selected from a nucleic acid assay, an antibody assay, and a cytometric assay.
  • an initial assay may comprise a cytometric assay, and a subsequent assay may comprise an assay selected from a nucleic acid assay, an antibody assay, and a general chemistry assay.
  • an initial assay comprises a cytometric assay
  • the cytometric assay may comprise the detection or measurement of a morphological characteristic of a cell.
  • a subsequent assay may comprise a cytometric assay for the detection or measurement of a characteristic of the sample; in embodiments, such a cytometric assay may comprise detection or measurement of a morphological characteristic of a ceil in the sample.
  • the methods may further comprise reporting the results of said initial assay of said biological sample. In embodiments, the results of an initial assay are reported prior to the performance of said subsequent assay. In embodiments of the methods disclosed herein, the methods may further comprise reporting the results of said subsequent assay. In embodiments of the methods disclosed herein, the methods may further comprise reporting the results of an initial assay and of a subsequent assay. In embodiments of the methods disclosed herein, the results of an initial assay and of a subsequent assay may be reported together.
  • the results of a subsequent assay are not reported if: i) said initial assay provides a negative result, and said negative result does not require performance of a subsequent assay, or ii) said initial assay provides a positive result, and said positive result does not require performance of a subsequent assay.
  • a small-volume biological sample may comprise a volume of less than about 200 microliters (uL), or of less than about 150 uL, or of less than about 100 , uL, or of less than about 75 ⁇ ,, or of less than about 50 ⁇ ,, or of less than about 40 ⁇ iL, or of less than about 30 jxL, or of less than about 20 ⁇ ., or of less than about 10 uL, or of less than about 5 ⁇ ,, or less.
  • uL microliters
  • a small-volume biological sample may comprise a sample selected from the group of biological sample types consisting of blood, serum, plasma, a throat swab, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid, tears, stool, mucus, sweat, earwax, oil, a glandular secretion, semen, vaginal fluid, interstitial fluids derived from tumorous tissue, ocular fluids, breath, hair, finger nails, skin, biopsy tissue, placental fluid, amniotic fluid, cord blood, lymphatic fluids, cavity fluids, sputum, pus, microbiota, meconium, breast milk, and other secretions or excretions.
  • biological sample types consisting of blood, serum, plasma, a throat swab, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid, tears, stool,
  • Applicant discloses herein methods of testing a small-volume biological sample in a de vice, wherein said small- volume biological sample has a volume of less than about 250 microliters ( , iiL), the methods comprising: placing said small-volume sample within the device; dividing said small-volume biological sample into at least a first and a second portion; performing, in said device on said first portion or aliquot thereof, a first assay for detecting or measuring an analyte or characteristic of the sample, whereby a first result is obtamed from the first portion, wherein said first result may comprise a negative result indicating that the presence of said analyte or characteristic is not detected or measured in the biological sample, or is def ected or measured at a normal level in the biological sample, or may comprise a positive result indicating that the presence of the analyte or characteristic is detected or measured in the biological sample, or is detected or measured at an abnormal level in the biological sample; performing, in said device on said second portion or aliquot
  • the analyte or characteristic to be detected or measured by a second assay may comprise a different analyte or characteristic than the analyte or characteristic detected or measured by a first assay.
  • the first assay may comprise an assay of an assay type selected from the group of assay types consisting of antibody-based assays, nucleic acid assays, general chemistry assays, and cytometric assays
  • said second assay comprises a different type of assay than said first assay.
  • a cytometric assay may comprise the detection or measurement of a morphological characteristic of a cell.
  • a first portion the small-volume sample may be diluted prior to performance of a first assay, or a second portion the small-volume sample may be diluted prior to performance of a second assay, or both.
  • a small-volume biological sample may comprise a sample selected from the group of biol ogical sample types consisting of blood, serum, plasma, a throat swa b, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid, tears, stool, mucus, sweat, earwax, oil, a glandular secretion, semen, vaginal fluid, interstitial fluids derived from tumorous tissue, ocular fluids, breath, hair, finger nails, skin, biopsy tissue, placental fluid, amniotic fluid, cord blood, lymphatic fluids, cavity fluids, sputum, pus, microbiota, meconium, breast
  • Applicant further discloses devices and sy stems suitable for the practice of the methods disclosed herein. Accordingly, Applicant discloses devices for testing a biological sample, wherein the device is configured to perform an initial assay and a subsequent assay according to the method disclosed herein (e.g., according to the methods disclosed above, and according to other methods disclosed herein).
  • a device for testing a biological sample may be configured to perform antibody-based assays, nucleic acid-based assays, general chemistry assays, and cytometric assays.
  • a device for testing a biological sample may be configured to test a blood sample.
  • a system as disclosed herein may comprise a device for testing a biological sample, e.g., a device as disclosed herein.
  • a system as disclosed herein may comprise a device for testing a biological sample that is configured to perform antibody-based assays, nucleic acid- based assays, general chemistry assays, and cytometric assays (including, e.g., detection, identification, or measurement of a morphological characteristic of a cell).
  • a system as disclosed herein may comprise a device for testing a biological sample that is configured to test a blood sample.
  • Applicant discloses herein methods of testing a small- volume biological sample in a device, wherein said small-volume biological sample has a volume of less than about 250 microliters (uL), the method comprising: placing said small-volume biological sample and a cartridge comprising reagents within the device; dividing ihe sample into at least a first and a second portion; retaining said second portion on said cartridge within the device for use in a subsequent assay; performing, in the device on said first portion or aliquot thereof, an initial assay for detecting, identifying, or measuring an analyte or characteristic of the sample, wherein said initial assay is performed using only reagents from said cartridge, whereby an initial result is obtained from the first portion, wherein: i) said initial result may comprise a negative result indicating (hat the presence of said analyte or characteristic is not detected, identified, or measured in the sample, or is detected, identified, or measured at a normal level in the sample, or ii) said initial result may comprise a
  • the small- volume biological sample has a volume of less than about 200 microliters i ui . s.
  • microliters or less than about 150 microliters (uL), or less than about 100 microliters (uL), or less than about 75 microliters (uL), or less than about 50 microliters (uL), or less than about 40 microliters (uL), or less than about 30 microliters ( , uL), or less than about 25 microliters ( ⁇ -.), or less than about 15 microliters (uL), or less than about i 0 microliters ( ⁇ ,), or less than about 5 microliters (uL), or less.
  • the sample comprises a sample selected from the group of biological sampl e types consisting of blood, serum, plasma, a throat swab, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid, tears, stool, mucus, sweat, earwax, oil, a glandular secretion, semen, vaginal fluid, interstitial fluids derived from tumorous tissue, ocular fluids, breath, hair, finger nails, skin, biopsy tissue, placental fluid, amniotic fluid, cord blood, lymphatic fluids, cavity fluids, sputum, pus, microbiota, meconium, breast milk, and other secretions or excretions.
  • biological sampl e types consisting of blood, serum, plasma, a throat swab, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid,
  • the sample is provided in a sample container that is disposed on said cartridge during placement of the sample within the device, and, following dividing the sample into at least a first and a second portion, the second portion is retained in the sample container (on the cartridge within the device) for use in the subsequent assay.
  • the sample is provided in a sample container that is disposed on the cartridge during placement of the sample within the device, and, fol lowing dividing the sample into at least a first and a second portion, the second portion is retained in a second container that is disposed on the cartridge within the device.
  • the second container is a dedicated container designated for holding the second portion on the cartridge prior to the performance of the subsequent assay.
  • the second container is a dedicated container designated to contain the second portion during the performance of the subsequent assay.
  • the automatic performance of the subsequent assay is begun within a short amount of time after placing the sample within the device, wherein the short amount of time is about three hours or less, or about two hours or less, or about one hour or less, or about 45 minutes or less, or about 30 minutes or less, or about 20 minutes or less, or about 10 minutes or less, or about 5 minutes or less, or is a shorter amount of time.
  • the automatic performance of the subsequent assay is completed within a short amount of time after placing the sample within the device, wherein the short amount of time is about three hours or less, or about two hours or less, or about one hour or less, or about 45 minutes or less, or about 30 minutes or less, or about 20 minutes or less, or about 10 minutes or less, or about 5 minutes or less, or is a shorter amount of time.
  • the cartridge contains all consumables used in the performance of the initial assay and of the subsequent assay.
  • the fsrst portion of the sample is diluted by at l east about 10-fold prior to, or during, the performance of the initial assay, or the second portion of the sample is diluted by at least about 10-fold prior to, or during, the automatic performance of the subsequent assay, or bo th.
  • the volume of the diluted portion is less than about 500 microliters ( ⁇ .), or is less than about 400 ⁇ _ ⁇ , or is less than about 300 uL, or is less than about 250 ⁇ ., or is less than about 200 ⁇ ., or is less than about 100 uL, or is less than about 50 pL, or is less than about 40 uL, or is less than about 30 ⁇ L ⁇ , or is less than about 20 microliters ⁇ ,.
  • the first portion of the sample is diluted by at least about 30- fold prior to, or during, the performance of the initial assay, or the second portion of the sample is diluted by at least about 30-fold prior to, or during, the automatic performance of the subsequent assay, or both.
  • the volume of the diluted portion is less than about 500 microliters ( ⁇ ⁇ ), or is less than about 400 ⁇ ,, or is less than about 300 , LiL, or is less than about 250 uL, or is less than about 200 ⁇ _, or is less than about 100 fiL, or is less than about 50 ⁇ , or is less than about 40 ⁇ ,, or is less than about 30 uL, or is less than about 20 microliters ⁇ .
  • the first portion of the sample is diluted by at least about 100-fold prior to, or during, the performance of the initial assay, or the second portion of the small-volume sample is diluted by at least about 100-fold prior to, or during, the automatic performance of the subsequent assay, or both.
  • the volume of the diluted portion is less than about 500 microliters ( ⁇ or is less than about 400 ⁇ ., or is less than about 300 ⁇ ., or is less than about 250 pL, or is less than about 200 ⁇ , or is less than about 100 ⁇ ,, or is less than about 50 ⁇ _,, or is less than about 40 ⁇ _, or is less than about 30 or is less than about 20 microliters ⁇ ,.
  • the initial assay comprises an assay of an assay type selected from the group of assay types consisting of antibody-based assays, nucleic acid assays, general chemistry assays, and cytometric assays.
  • the subsequent assay comprises a different type of assay than the initial assay.
  • cytometric assays may comprise the detection, identification, or measurement of a morphological characteristic of a cell.
  • the subsequent assay comprises an assay for detecting, identifying, or measuring: a) the same analyte or characteristic as the initial assay, wherein the subsequent assay is more sensitive for the detection, identification, or measurement of the analyte or characteristic than the initial assay, or b) a different analyte or characteristic than the analyte or characteristic detected, identified, or measured by the initial assay.
  • the initial assay comprises a nucleic acid assay
  • the subsequent assay comprises an assay selected from an antibody-based assay, a general chemistry assay, and a cytometric assay
  • the initial assay comprises an antibody assay
  • the subsequent assay comprises an assay selected from a nucleic acid assay, a general chemistry assay, and a cytometric assay
  • the initial assay comprises a general chemistry assay
  • the subsequent assay comprises an assay selected from a nucleic acid assay, an antibody assay, and a cytometric assay
  • the initial assay comprises a cytometric assay
  • the subsequent assay comprises an assay selected from a nucleic acid assay, an antibody assay, and a general chemistry assay.
  • the cytometric assay may comprise the detection, identification, or measurement of a morphological characteristic of a cell.
  • the initial assay may comprise the detection, identification, or measurement of an analyte
  • the subsequent assay may comprise a cytometric assay for the detection, identification, or measurement of a morphological characteristic of a cell.
  • the methods further comprise reporting the results of the initial assay of the sample.
  • the results of the initial assay are reported prior to the performance of the subsequent assay.
  • methods of testing a small-volume biological sample in a device comprise reporting the results of the initial assay of the sample and further comprise reporting the results of the subsequent assay.
  • the results of the initial assay and of the subsequent assay are reported together.
  • Applicant discloses herein methods of testing a small-volume biological sample in a device, wherein said sample has a volume of less than about 250 microliters (jjJL), the method comprising: placing the sample and a cartridge comprising reagents within the device; dividing the sample into at least a first and a second portion, and retaining at least said second portion on said cartridge prior to performing assays on said portions; performing, in said device on the first portion or aliquot thereof, a first assay for detecting, identifying, or measuring an analyte or characteristic of the sample using only reagents from said cartridge, whereby a first result is obtained from the first portion, wherein said first result may comprise a negative result indicating that the presence of said analyte or characteristic is not detected or measured in the sample, or is detected or measured at a normal level in the sample, or may comprise a positive result indicating that the presence of the analyte or characteristic is detected or measured in the sample, or is detected or measured at an abnormal
  • the methods comprise retaining both said first and said second portions on said cartridge prior to performing assays on said portions.
  • the small- volume biological sample has a volume of less than about 200 microliters (fiL), or less than about 150 microliters ( L), or less than about 100 microliters ( ⁇ -), or less than about 75 microliters ( ⁇ ,), or less than about 50 microliters (jjJL), or less than about 40 microliters (uL), or less than about 30 microliters (uL), or less than about 2.5 microliters (uL), or less ihan about 15 microliters ⁇ , ⁇ . ⁇ . or less than about 10 microliters (fiL), or less than about 5 microliters (uL), or less.
  • the sample comprises a sample selected from the group of biological sample types consisting of blood, serum, plasma, a throat swab, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid, tears, stool, mucus, sweat, earwax, oil, a glandular secretion, semen, vaginal fluid, interstitial fluids derived from tumorous tissue, ocular fluids, breath, hair, finger nails, skin, biopsy tissue, placental fluid, amniotic fluid, cord blood, lymphatic fluids, cavity fluids, sputum, pus, microbiota, meconium, breast milk, and other secretions or excretions.
  • biological sample types consisting of blood, serum, plasma, a throat swab, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid, tears, stool, mucus, sweat,
  • the second assay comprises an assay for detecting, identifying, or measuring: a) the same analyte or characteristic as the first assay, and wherein the second assay is more sensitive for the detection or measurement of the analyte or characteristic than the first assay, or b) a different analyte or characteristic than the analyte or characteristic detected, identified, or measured by the first assay.
  • the first assay comprises an assay of an a ssay type selected from the group of assay types consisting of antibody-based assays, nucleic acid assays, general chemistry assays, and cytometric assays.
  • a cytometric assay may comprise the detection, identification, or measurement of a morphological characteristic of a cell.
  • the second assay comprises a different type of assay than the first assay.
  • the cartridge contains all consumables used in the performance of the first assay and of the second assay.
  • the first portion of the sample is diluted prior to, or during, the performance of the first assay, or wherein the second portion of the sample is diluted prior to, or during, the perfonnance of the second assay, or both.
  • the first portion of the sample is diluted by at least about 10-fold prior to, or during, the performance of the initial assay, or the second portion of the sample is diluted by at least about 10-fold prior to, or during, the automatic performance of the subsequent assay, or both.
  • the volume of the diluted portion is less than about 500 microliters (uL), or is less than about 400 uL, or is less than about 300 iL, or is less than about 250 uL, or is less than about 200 ⁇ , or is less than about 100 , nL, or is less than about 50 ⁇ ,, or is less than about 40 iL, or is less than about 30 ⁇ ,, or is less than about 20 microliters iL.
  • the first portion of the sample is diluted by at least about 30- fold prior to, or during, the performance of the initial assay, or the second portion of the sample is diluted by at least about 30-fold prior to, or during, the automatic performance of the subsequent assay, or both.
  • the volume of the diluted portion is less than about 500 microliters ( ⁇ ⁇ ), or is less than about 400 ⁇ ,, or is less than about 300 ,i!.L, or is less than about 250 ⁇ , or is less than about 200 L, or is less than about 100 uL, or is less than about 50 uL, or is less than about 40 ⁇ L ⁇ , or is less than about 30 uL, or is Jess than about 20 microliters ⁇ ,.
  • the first portion of the sample is diluted by at least about 100-fold prior to, or during, the performance of the initial assay, or the second portion of the sample is diluted by at least about 100- fold prior to, or during, the automatic performance of the subsequent assay, or both.
  • the volume of the diluted portion is less than about 500 microliters (uL), or is less than about 400 uL, or is less than about 300 uL, or is less than about 250 ⁇ _, or is less than about 200 ⁇ ,, or is less than about 100 uL, or is less than about 50 uL, or is Jess than about 40 ⁇ ,, or is less than about 30 ⁇ ,, or is less than about 20 microliters ⁇ ...
  • Applicants disclose herein devices for testing a small-volume biological sample wherein said devices are configured to perform an initial assay and a subsequent assay according to the methods disclosed herein.
  • a device disclosed herein is configured to perform antibody-based assays, nucleic acid-based assays, general chemistry assays, and cytometric assays.
  • a device disclosed herein is configured to test the small-volume biological sample, wherein the sample is selected from the group of biological sample types consisting of blood, serum, plasma, a throat swab, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid, tears, stool, mucus, sweat, earwax, oil, a glandular secretion, semen, vaginal fluid, interstitial fluids derived from tumorous tissue, ocular fluids, breath, hair, finger nails, skin, biopsy tissue, placental fluid, amniotic fluid, cord blood, lymphatic fluids, cavity fluids, sputum, pus, microbiota, meconium, breast milk, and other secretions or excretions.
  • a device disclosed herein is configured to test a blood sample.
  • Applicants disclose herein devices for testing a small-volume biological sample wherein said systems comprise a device as disclosed herein.
  • a system disclosed herein is configured to perform antibody-based assays, nucleic acid-based assays, general chemistry assays, and cytometric assays.
  • the systems comprise a device as disclosed herein, wherein the device is configured to test a small- volume biological sample, where the small- volume biological sample is a sample as disclosed herein.
  • the systems comprise a device as disclosed herein, wherein the device is configured to test a blood sample.
  • the assays and methods disclosed herein may be performed on a device, or on a system, for processing a sample.
  • the assays and methods disclosed herein can be readily incorporated into and used in device for processing a sample, or a system for processing a sample, which may be an automated assay device, or may be an automated assay system.
  • a device and such a system, may be useful for the practice of the methods disclosed herein.
  • a device may be useful for receiving a sample.
  • a device may be useful for preparing, or for processing a sample.
  • a device may be useful for performing an assay on a sample.
  • a device may be useful for obtaining data from a sample.
  • a device may be useful for transmitting data obtained from a sample.
  • a device may be useful for disposing of a sample following processing or assaying of a sample,
  • a device may be part of a system, a component of which may be an automated assay device.
  • a device may be an automated assay device.
  • An automated assay device may ⁇ be configured to facilitate collection of a sample, prepare a sample for a clinical test, or effect a chemical reaction with one or more reagents or other chemical or physical processing, as disclosed herein.
  • An automated assay device may be configured to obtain data from a sample.
  • An automated assay device may be configured to transmit data obtained from a sample.
  • An automated assay device may be configured to analyze data from a sample.
  • An automated assay device may be configured to communicate with another device, or a laboratory, or an individual affiliated with a laboratory, to analyze data obtained from a sample.
  • a automated assay device may be configured to accept a sample from a subject, either directly or indirectly.
  • a sample may be, for example, a blood sample (e.g., a sample obtained from a tmgerstick, or from venipuncture, or an arterial blood sample), a urine sample, a biopsy sample, a tissue slice, stool sample, or other biological sample; a water sample, a soil sample, a food sample, an air sample; or other sample.
  • a blood sample may comprise, e.g., whole blood, plasma, or serum.
  • An automated assay device may receive a sample from the subject through a housing of the device. The sample collection may occur at a sample collection site, or elsewhere. The sample may be provided to the device at a sample collection site.
  • samples may include various fluid or solid samples.
  • the sample can be a bodily fluid sample from the subject
  • the sample can be an aqueous or gaseous sample.
  • solid or semi-solid samples can be provided.
  • the sample can include tissues and/or cells collected from the subject.
  • the sample can be a biological sample.
  • sample volumes that may be obtained from a subject include, but are not limited to, volumes of about: 10 mL or less, 5 lnL or less, 3 mL or less, 1 microliter ( ⁇ _, also "uL” herein) or less, 500 ⁇ ., or less, 300 aL or less, 250 ⁇ , or less, 200 ⁇ , or less, 170 ⁇ , or less, 150 ⁇ _ or less, 125 ⁇ L or less, 100 ⁇ , or less, 75 ⁇ .
  • ⁇ _ also "uL” herein
  • a biological sample may have a volume of: about 250 jxL or less; or about 200 ⁇ , or less; or about 150 ⁇ , or less; or about 100 JJL or less; or about 75 ⁇ , or less; or about 50 ⁇ . or less; or about 40 uL or less; or about 30 ⁇ . or less; or about 20 ⁇ , or less; or about 10 ⁇ , or less; or about 5 or less; or about 4 ⁇ . or less; or about 3 or less; or about 2 ⁇ ., or less; or about 1 ⁇ ,, or less.
  • the amount of sample may be about a drop of a sample.
  • the amount of sample may be the amount collected from a pricked finger or fingerstick.
  • the amount of sample may be the amount collected from a microneedle or a venous draw. Any volume, including those described herein, may be provided to the device.
  • a biological sample may include cells.
  • a sample such as a biological sample
  • a biological sample may be contained within a sample collection device, such as a vial, a tube, a jar, a specimen bottle, or other container or device suitable for holding or transporting a biological sample.
  • a sample collection device may be or comprise a small volume container.
  • a sample collection device may be or comprise a small volume container having a holding capacity of about 2.50 ⁇ , or less, or about 200 pL or less, or about 150 ⁇ . or less, or about 100 ⁇ -L or less, or about 50 ⁇ .
  • a sample collection device may be or comprise a nanotainer *M vessel.
  • the biological sample delivered by a cartridge is a processed biological sample.
  • the biological sample delivered by a cartridge is a processed biological sample, and, following delivery of the processed biological sample, the processed biological sample is mixed with a reageni within an automated assay device or system.
  • the biological sample delivered by a cartridge is a processed biological sample, and the processed biological sample is mixed with a reagent prior to detection of the presence or absence of a target analyte or characteristic in the processed biological sample within an automated assay device or system.
  • the biological sample delivered by a cartridge is a diluted biological sample.
  • the biological sample delivered by a cartridge is a diluted biological sample, and, following deliver of the diluted bioiogicai sample, the diluted bioiogicai sample is mixed with a reagent within an automated assay device or system.
  • the biological sample delivered by a cartridge is a diluted biological sample, and the diluted biological sample is mixed with a reagent prior to detection of the presence or absence of a target analyte or characteristic in the diluted biological sample within an automated assay device or system.
  • the biological sample delivered by a cartridge is an undiluted bioiogicai sample.
  • ihe biological sample delivered by a cartridge is an undiluted bioiogicai sample and the biological sample is diluted within an automated assay device or system.
  • the biological sample delivered by a cartridge is an undiluted bioiogicai sample and the biological sample is diluted within an automated assay device or system prior to analysis of the biological sample.
  • the biological sample delivered by a cartridge is an undiluted biological sample and the first step applied to the biological sample within the automated assay device or system is a dilution step. In embodiments, such a first step is performed within an automated assay device or system.
  • the biological sample delivered by a cartridge is an undiluted biological sample, and the biological sample is diluted and then mixed with a reagent prior to detection of the presence or absence of a target analyte or characteristic in the diluted bioiogicai sample within an automated assay device or system.
  • an assay may utilize a diluted sample (e.g., a diluted bioiogicai sample).
  • a diluted sample e.g., a diluted bioiogicai sample.
  • an assay may include instructions, or a protocol, for the dilution of a biological sample, in embodiments, such dilution may provide a diluted sample diluted by at least about 10-fold as compared to the original sample; or may provide a diluted sample diluted by at least about 20- fold as compared to the original sample: or may provide a diluted sample diluted by at least about 30-fold as compared to the original sample; may provide a diluted sample diluted by at least about 50-fold as compared to the original sample; or may provide a diluted sample diluted by at least about 100-fold as compared to the original sample; or may provide a diluted sample diluted by at least about 200-fold as compared to the
  • a cartridge may contain a biological sample; a reagent unit; and an assay unit.
  • a cartridge may contain a biological sample: a reagent unit: an assay unit; and a calibration unit.
  • a cartridge may contain a biological sample; a reagent unit; an assay unit; a calibration unit; and a pipette.
  • a cartridge may contain a biological sample; a reagent unit; an assay unit; a calibration unit; and a vessel.
  • a cartridge may contain a biological sample; a reagent unit; an assay unit; a calibration unit a pipette; and a vessel.
  • a cartridge may comprise an identifying mark, or label, or other identification.
  • a cartridge may contain one or more biological samples; one or more reagent units; one or more assay units; one or more calibration units; one or more pipettes; and may contain combinations thereof.
  • a cartridge may include a vessel, or a plurality of vessels, in addition to biological samples, reagent units, assa units, calibration units, pipettes, and combinations thereof.
  • a cartridge may lack a biological sample; for example, in embodiments, a biological sample may be provided to an automated assay device or system separately from a cartridge, where the cartridge lacking a biological sample may contain one or more reagent units, one or more assay units, one or more calibration units, one or more pipettes, one or more vessels, and combinations thereof.
  • an automated assay device may be configured to accept or hold a cartridge.
  • an automated assay device may comprise a cartridge.
  • the cartridge may be removable from the automated assay device.
  • a sample may be provided to the cartridge of the automated assay device.
  • a sample may be prtwided to another portion of an automated assay device.
  • the cartridge and/or device may comprise a sample collection unit that may be configured to accept a sample.
  • a cartridge may include a sample, and may include reagents for use in processing or testing a sample, disposables for use in processing or testing a sample, or other materials. Following placemen! of a cartridge on, or insertion of a cartridge into, an automated assay device, one or more components of the cartridge may be brought into fluid communication with other components of the automated assay device. For example, if a sample is collected at a cartridge, the sample may be transferred to other portions of the automated assay de vice. Similarly, if one or more reagents are provided on a cartridge, the reagents may be transferred to other portions of the automated assay device, or other components of the automated assay device may be brought to the reagents. In some embodiments, the reagents or components of a cartridge may remain on-board the cartridge. In some embodiments, no fluidics are included that require tubing or that require maintenance (e.g., manual or automated maintenance).
  • a cartridge may include a biological sample, or may include two or more biological samples.
  • a biological sample is from a subject, and multiple biological samples may be from a single subject. In alternative embodiments, multiple biological samples may be from multiple subjects.
  • the device may include an automated lancing cartridge.
  • the cartridge may be disposable.
  • One or more disposable component may be used to collect a sample from a subject.
  • the disposable component may provide the sample to a non-disposable device.
  • the disposable component may be the automated assay device.
  • a cartridge may include a reagent or a plurality of reagents, and may be configured to allow delivery of said reagent or reagents to said de vice.
  • a cartridge may be configured to deliver a biological sample and a reagent to the device.
  • a cartridge may be configured to deliver a plurality of biological samples and a reagent to the device.
  • a cartridge may be configured to deliver a biological sample and a plurality of reagents to the device.
  • a cartridge may be configured to deliver a plurality of biological samples and a plurality of reagents to the device.
  • Identification information may include subject identifying information, information based on signals generated related to the sample, information based on signals generated related to reactions performed with the sample, information based on signals detected related to the sample, information based on signals detected related to reactions performed with the sample, device identification information, cartridge identification information, component identifying information, and other information transmitted from the device,
  • a sample or reagent carried by a cartridge may be transferred to a device, such as an automated assay device.
  • a sample or reagent may be transferred within a device. Such transfer of sample or reagent may be accomplished without providing a continuous fluid pathway from cartridge to device. Such transfer of sample or reagent may be accomplished without providing a continuous fluid pathway within a device.
  • such transfer of sample or reagent may be accomplished by a fluid handling system (e.g., a pipette); for example, a sample, reagent, or aliquot thereof may be aspirated into an open-tipped transfer component, such as a pipette tip, which may be operably connected to a fluid handling system which transfers the tip, with the sample, reagent, or aliquot thereof contained within the tip, to a location on or within the automated assay device.
  • the sample, reagent or aliquot thereof can be deposited at a location on or within the automated assay device.
  • Sample and reagent, or multiple reagents may be mixed using a fluid handling system in a similar manner.
  • One or more components of ihe cartridge may be transferred in an automated fashion to other portions of the automated assay device, and vice versa,
  • a fluid handling system may be used to transport and deliver a sample solution to a vessel, and to fill a vessel (either partially or fully) with a sample solution.
  • a fluid handling system comprises a pipette.
  • a pipette may be configured to accept a pipette tip, e.g., to mount and transport a pipette tip attached to the pipette.
  • a pipette comprises a nozzle configured to accept a pipette tip, A.
  • pipette may be configured to aspirate a fluid, such as a sample solution, into a pipette tip attached to the pipette (e.g., a pipette tip which is attached to a nozzle of the pipette).
  • a pipette may be configured to dispense a fluid, such as a sample solution, from a pipette tip attached to the pipette (e.g., to a nozzle of the pipette).
  • a pipette may be configured to transmit force to a surface or component of a device.
  • a pipette nozzle may contact a surface or component of a device, effective to transmit force to thai surface or component.
  • a pipette nozzle may contact a mating recess of a vessel, and, in embodiments, may engage a mating recess of a vessel.
  • two, or more pipette nozzles may contact mating recesses of a vessel and, in embodiments, may engage mating recesses of a vessel
  • a pipette of a fluid handling system may be movable, and is preferably movable in at least two, and more preferably in three dimensions (e.g., is movable in one, two, or all three of horizontally, laterally, and vertically).
  • a device such as an automated assay device, may have a fluid handling system.
  • a fluid handling system may perform, or may aid in performing, transport, dilution, extraction, aliquorting, mixing, and other actions with a fluid, such as a sample.
  • a fluid handling system may be contained within a device housing.
  • a fluid handling system may permit the collection, delivery, processing and/or transport of a fluid, dissolution of dry reagents, mixing of liquid and/or dry reagents with a liquid, as well as collection, delivery, processing and/or transport of non-fluidic components, samples, or materials.
  • the fluid may be a sample, a reagent, diluent, wash, dye, or any other fluid that may be used by the device, and may include, but not limited to, homogenous fluids, different liquids, emulsions, suspensions, and other fluids.
  • a fluid handling system including without limitation a pipette, may also be used to transport vessels (with or without fluid contained therein) around the device.
  • the fluid handling system may dispense or aspirate a fluid.
  • the sample may include one or more paniculate or solid matter floating within a fluid.
  • a fluid handling system may comprise a pipette, pipette tip, syringe, capillary, or other component.
  • the fluid handling system may have portion with an interior surface and an exterior surface and an open end.
  • the fluid handling system may comprise a pipette, which may include a pipette body and a pipette nozzle, and may comprise a pipette tip.
  • a pipette tip may or may not be removable from a pipette nozzle.
  • a fluid handling system may use a pipette mated with a pipette tip; a pipette tip may be disposable, A tip may form a fluid- tight seal when mated with a. pipette.
  • a pipette tip may be used once, twice, or more times.
  • a fluid handling system may use a pipette or similar device, with or without a pipette tip, to aspirate, dispense, mix, transport, or otherwise handle the fluid.
  • the fl id may be dispensed from the fluid handling sy tem when desired.
  • the fluid may be contained within a pipette tip prior to being dispensed, e.g., from an orifice in the pipette tip. In embodiments, or instances during use, all of the fluid may be dispensed; in other embodiments, or instances during use, a portion of the fluid within a tip may be dispensed.
  • a pipette may selectively aspirate a fluid.
  • the pipette may aspirate a selected amount of fluid.
  • the pipette may be capable of actuating stirring mechanisms to mix the fluid within the tip or within a vessel.
  • the pipette may incorporate tips or vessels creating continuous flow loops for mixing, including of materials or reagents that are in non-liquid form.
  • a pipette tip may also facilitate mixture by metered delivery of multiple fluids simultaneously or in sequence, such as in two-part substrate reactions.
  • the fluid handling system may include one or more fluidically isolated or hydraulicaliv independent units.
  • the flu d handling system may include one, two, or more pipette tips.
  • the pipette tips may be configured to accept and confine a fluid.
  • the tips may be fluidically isolated from or hydrauiically independent of one another.
  • the fluid contained within each tip may be fluidically isolated or hydrauiically independent from one fluids in other tips and from other fluids within the device.
  • the fluidically isolated or hydrauiically independent units may be movable relative to other portions of the device and/or one another.
  • the fluidically isolated or hydrauiically independent units may be individually movable.
  • a fluid handling system may comprise one or more base or support.
  • a base or support may support one or more pipette or pipette units.
  • a base or support may connect one or more pipettes of the fluid handling system to one another,
  • An automated assay device may be configured to perform processing steps or actions on a sample obtained from a subject.
  • Sample processing may include sample preparation, including, e.g., sample dilution, division of a sample into aliquots, extraction, contact with a reagent, filtration, separation, centrifugation, or other preparatory or processing action or step.
  • An automated assay device may be configured to perform one or more sample preparation action or step on the sample.
  • a sample may be prepared for a chemical reaction and/or physical processing step.
  • sample preparation action or step may include one or more of the following: centrifugation, separation, filtration, dilution, enriching, purification, precipitation, incubation, pipetting, transport, chromatography, cell lysis, cytometry, pulverization, grinding, activation, uiirasonication, micro column processing, processing with magnetic beads, processing with nanoparticles, or other sample preparation action or steps.
  • sample preparation may include one or more step to separate blood into serum and/or particulate fractions, or to separate any other sample into various components.
  • Sample preparation may include one or more step to dilute and/or concentrate a sample, such as a blood sample, or other biological samples.
  • Sample preparation may include adding an anti-coagulant or other ingredients to a sample.
  • Sample preparation may also include purification of a sample.
  • ail sample processing, preparation, or assay actions or steps are performed by a single device.
  • all sample processing, preparation, or assay actions or steps are performed within a housing of a single device.
  • most sample processing, preparation, or assay actions or steps are performed by a single device, and may be performed within a housing of a single device, in embodiments, many sample processing, preparation, or assay- actions or steps are performed by a single device, and may be performed within a housing of a single device.
  • sample processing, preparation, or assay actions or steps may be performed by more than one device.
  • An automated assay device may be configured to run one or more assay on a sample, and to obtain data from the sample.
  • An assay may include one or more physical or chemical treatments, and may include running one or more chemical or physical reactions.
  • An automated assay device may be configured to perform one, two or more assays on a small sample of bodily fluid. One or more chemical reaction may take place on a sample having a volume, as described elsewhere herein.
  • the sample collection unit is configured to receive a volume of the bodily fluid sample equivalent to a single drop or less of sample or interstitial fluid
  • the volume of a sample may be a small volume, where a small volume may be a volume that is less than about 1000 ⁇ ,, or less than about 500 fjL, or less than about 250 fjL, or less than about 150 ⁇ _, or less than about 100 ⁇ _, or less than about 75 ⁇ 3_, or less than about 50 ⁇ £, or less than about 40 uL, or less than about 20 uL, or less than about 10 ⁇ ,, or other small volume, in embodiments, all sample assay actions or steps are performed by a single device. In embodiments, sample processing, preparation, or assay actions or steps may be performed by more than one device.
  • An automated assay device may be configured to perform a plurality of assays on a sample.
  • an automated assay device may be configured to perform a plurality of assays on a single sample.
  • a plurality of assay s may be run simultaneously; may be run sequentially; or some assays may be rim simultaneously while others are rim sequentially.
  • One or more control assays and/or calibrators can also be incorporated into the device; control assays and assay on calibrators may be performed simultaneously with assays performed on a sample, or may be performed before or after assays performed on a sample, or any combination thereof.
  • sample assay actions or steps, of a plurality of assays are performed by a single device, and may be performed within a housing of a single device, in embodiments, sample processing, preparation, or assay actions or steps may be performed by more than one device.
  • devices, and systems and methods comprising or using such devices may comprise a detector configured to detect analyte in a sample.
  • a detector may be, for example, an optical detector, such as a spectrophotometer, a photomultiplier, a charge- coupled device, a camera, or other device or system configured to detect a light-based signal indicative of the presence of a analyte.
  • a detector may be configured to, or be effective to, detect a signal comprising chemilummescence, luminescence, fluorescence, absorbance, transmittance, turbidity, a color change, or other change in light, whether emitted, transmitted, or absorbed, effective to signal the presence of a analyte in a sample.
  • a detector may comprise an electrochemical detector, or a temperature sensor, or a pH sensor, or a radiation sensor, or an ion-sensitive electrode, or other sensor capable of detecting the presence of a analyte in a sample.
  • Methods for detecting analyte include assays for detecting nucleic acids (e.g., DMA or RNA), assays for detecting peptides and proteins (including glycoproteins), assays for detecting other pathogen-related molecules, complement fixation assays,
  • nucleic acid assays include polymerase chain reaction (PGR) methods (including quantitative PGR (qPCR), reverse -transcriptase PCD (RT-PCR), "realtime” PGR, one-step PGR., two-step PGR, and other methods known, in the art.
  • PGR polymerase chain reaction
  • Methods for detecting peptides and proteins include "antibody-based assays” such as, e.g., enzyme immunoassays such as Enzyme-Linked Immunosorbent Assays (ELISAs) and other assays utilizing antibodies or antibody fragments, complement-based reactions, measurement of absorbance of ultraviolet or other frequency of light, assays utilizing specific receptor- ligand interactions, and other assays known in the art.
  • Assays for detecting other pathogen-related molecules include assay s for bacterial sugars and lipids (e.g., bacterial lipopolysaccharide (LPS)), and other assays known in the art.
  • a detector for use with such assays may be an optical detector, a pH detector, an electrochemical detector, a temperature sensor, an ion- sensitive electrode, a radiation detector, or other detector.
  • An automated assay device may be configured to detect one or more signals relating to the sample.
  • An automated assay device may be configured to identify one or more properties of the sample. For instance, the automated assay device may be configured to detect the presence or concentration of one analyte or a plurality of analytes or a disease condition in the sample (e.g., in or through a bodily fluid, secretion, tissue, or other sample).
  • the automated assay device may be configured to detect a signal or signals that may be analyzed to detect the presence or concentration of one or more analytes (which may be indicative of a disease condition) or a disease condition in the sample.
  • the signals may be analyzed on board the device, or at another location. Running a clinical test may or may not include any analy sis or comparison of data collected.
  • a chemical reaction or other processing step may be performed, with or without the sample.
  • steps, tests, or assays thai may be prepared or run by the device may include, but are not limited to immunoassay, nucleic acid assay, receptor-based assay, cytometric assay, colorimetric assay, enzymatic assay, electrophoretic assay, electrochemical assay, spectroscopic assay, chromatographic assay, microscopic assay, topographic assay, calorimetric assay, turbidmetric assay, agglutination assay, radioisotope assay, viscometric assay, coagulation assay, clotting time assay, protein synthesis assay, histological assay, culture assay, osmolality assay, and/or other types of assays,
  • Steps, tests, or assays that may be prepared or run by the device may include imaging, including microscopy, cytometry, and other techniques preparing or utilizing images. Steps, tests, or assays that may be prepared or run by the device may further include an assessment of histology, morphology, kinematics, dynamics, and/ or state of a sample, which may include such assessment for cells,
  • a device may be capable of performing all on-board steps (e.g., steps or actions performed by a single device) prior to the performance of a reflex test in a short amount of time.
  • a device may be capable of performing all on-board steps on a single sample in a short amount of time prior to the performance of a reflex test.
  • the amount of time between the time of sample collection from a subject to the time of transmitting data and/or analysis prior to the performance of a reflex test may be a short amount of time.
  • the amount of time between the time of accepting or placing a sample within the device to the time of transmitting data and/or analysis from the device regarding such a sample may depend on the type or number of steps, tests, or assays performed on the sample.
  • the amount of time between the time of accepting or placing a sample within the device to the time of beginning to perform a subsequent assay may be a short amount of time.
  • the amount of time between the time of accepting or placing a sample within the device to the time of obtaining a result from the performance of a subsequent assay may be a short amount of time.
  • the amount of time between the time of accepting or placing a sample within the device to transmitting data and/or analysis from the device regarding such a sample, prior to the performance of a reflex test may be a short amount of time.
  • a short amount of time may comprise about 3 hours or less, about 2.
  • a device may be configured to prepare a sample for disposal, or to dispose of a sample, such as a biological sample, following processing or assaying of a sample.
  • an automated assay device may be configured to transmit data obtained from a sample.
  • an automated assay device may be configured to communicate over a network.
  • An automated assay device may include a communication module that may interface with the network.
  • An automated assay device may be connected to the network via a wired connection or wirelessly.
  • the network may be a local area network (LAN) or a wide area network (WAN) such as the Internet.
  • the network may be a personal area network.
  • the network may include the cloud.
  • the automated assay device may be connected to the network without requiring an intermediary device, or an intermediary device may be required to connect an automated assay device to a network.
  • An automated assay device may communicate over a network with another device, which may be any type of networked device, including but not limited to a personal computer, server computer, or laptop computer; personal digital assistants (PDAs) such as a Windows CE device; phones such as cellular phones, smartphones (e.g., iPhone, Android, Blackberry, etc.), or location -aware portable phones (such as GPS); a roaming device, such as a network-connected roaming device; a wireless device such as a wireless email device or other device capable of communicating wireless with a computer network; or any other type of network device that may communicate possibly o ver a network and handle electronic transactions.
  • PDAs personal digital assistants
  • a roaming device such as a network-connected roaming device
  • a wireless device such as a wireless email device or other device capable of communicating wireless with a computer network
  • any other type of network device that may communicate possibly o ver a network and handle electronic transactions.
  • Such communication may include providing data to a cloud computing infrastructure or any other type of data
  • An automated assay device may provide data regarding a sample to, e.g., a health care professional, a health care professional location, such as a laboratory, or an affiliate thereof.
  • a laboratory, health care professional, or subject may have a network device able to receive or access data provided by the automated assay device.
  • An automated assay device may be configured to provide data regarding a sample to a database.
  • An automated assay device may be configured to provide data regarding a sample to an electronic medical records system, to a laboratory information system, to a laboratory automation system, or other system or software.
  • An automated assay device may provide data in the form of a report.
  • devices, and systems and methods comprising or using such devices may comprise a controller.
  • a controller may comprise a processor.
  • a controller may be connected to, and may control the operation of, components of a device; such components are typically disposed within a housing of the device.
  • a controller may control the operation of a fluid handling system.
  • a controller may control the operation of a detector, m embodiments, a controller may control the operation of any component or unit of the device.
  • Other components may include, for example, a camera, a chemistry assay unit, a nucleic acid assay unit, a heating unit, a communication unit, a protein chemistry unit, or other component or unit.
  • a controller may control the operation of one or more components of a device according to a protocol.
  • a protocol by which a controller controls the operation of any one or more component or unit of a device may be preprogrammed, e.g., may be resident on the device.
  • a protocol by which a controller controls the operation of any one or more component or unit of a device may be obtained from another device, or from a user, or from a laboratory, or from a network, or from the cloud.
  • a protocol by which a controller controls the operation of any one or more component or unit of a device may be updated, or may be updatable, according to information or instructions from another device, or from a user, or from a laboratory, or from a network, or from the cloud.
  • a device may receive information, or instructions, or updates, or protocols, via a user interface.
  • a device may receive information, or instructions, or updates, or protocols, via a communication assembly.
  • devices, and systems and methods comprising or using such devices may comprise a display effective to provide a user with information regarding the operation of the device, information regarding the progress of an assay performed by the device, or information regarding the results of an assay performed by the device.
  • a display may comprise a visual display, or may comprise a printed display, or may comprise an audio signal, which may include an audio signal understandable as speech by a user, or may comprise any combination or ail of such displays.
  • a display may comprise a user interface.
  • a device may receive, e.g., information, commands, protocols, or other input.
  • a user interface may communicate a request for a second, or subsequent, biological sample.
  • a user interface may communicate instructions regarding obtaining a second, or subsequent, biological sample.
  • devices, and systems and methods comprising or using such devices may comprise a communication assembly effective to communicate with one or more of a user, another device, a laboratory, a network, the cloud, or other communication target.
  • a communication assembly may provide a communication target with information regarding the operation of the device, information regarding the progress of an assay performed by the de vice, or information regarding the results of an assay performed by the device.
  • a communication assembly may be configured to allow a device to receive, e.g., information, commands, protocols, or other input from an outside source, such as, e.g., a user, another device, a laboratory, a network, the cloud, or other communication source.
  • a protocol may include instructions regarding one or more of: preparation of a sample; preparation of a plurality of samples; performing a chemical reaction; performing a plurality of chemical reactions; sequence of performing a plurality of chemical reactions; performing a clinical test; performing a plurality of clinical tests;
  • protocol information may be changed according to transmitted data obtained from said biological sample within said housing of said device according to said protocol.
  • Order forms, methods of ordering clinical assays, and methods of performing clinical assays using such order forms are disclosed herein.
  • such order forms include listings of a plurality of assays which may be ordered, where these listings include prices for each of the assays which may be ordered.
  • methods for ordering clinical assays include marking order forms which include listings of a plurality of assays which may he ordered, where these listings include prices for each of the assays which may be ordered.
  • methods for performing clinical assays include ordering clinical assays using order forms which include listings of a plurality of assays which may be ordered, where these listings include prices for each of the assays which may be ordered, and performing the clinical assays. These clinical assays may be performed on samples obtained from subjects at a single sample acquisition session; such samples may be small volume samples, e.g., small volume samples of blood or urine.
  • Applicant discloses methods of ordering assays.
  • a method of ordering a clinical assay to be performed by a clinical testing facility on a sample obtained from a subject comprising: Providing an order form listing a plurality of assays which may be ordered, wherein the cost of each assay of said plurality of assays is provided on said order form; Marking the order form to indicate the name and other identifying information of said subject; Marking the order form to identify one or more selected assays of the plurality of assays that are ordered for performance; and Providing the marked order form to the clinical testing facility.
  • the sample is a small- volume biological sample having a volume of less than about 250 microliters (uL).
  • the sample is a blood sample or a urine sample.
  • the sample is provided in a sample container for performance of said assays by the clinical testing facility using an automated sample analysis device,
  • Further embodiments of these methods comprise ordering a plurality of assays comprising an initial assay and a subsequent assay, wherein the sample is provided in a sample container that is disposed on a cartridge during pla cement of the sample within an automated sample analysis device, wherein said sample is divided into portions, said portions comprising a first portion and at feast a second portion, and wherein said second portion is retained in said sample container disposed on the cartridge within the device for use in the subsequent assay.
  • the cartridge contains all consumables used in the performance of the initial assay and of the at least one subsequent assay.
  • the sample is a. small-volume biological sample having a volume of less than about 2.50 microliters (uL).
  • the sample is a blood sample or a urine sample.
  • the sample is provided in a sample container for performance of said assay s by the clinical testing facility using an automated sample ana lysis de v ice.
  • Embodiments of these methods comprise ordering a plurality of assays comprising an initial assay and a subsequent assay, wherein the sample is provided in a sample container that is disposed on a cartridge during placement of the sample within an automated sample analysis device, wherein said sample is divided into portions, said portions comprising a first portion and at least a second portion, and wherein said second portion is retained in said sample container disposed on the cartridge within the device for use in the subsequent assay.
  • the cartridge contains all consumables used in the performance of the initial a ssay and of the at least one subsequent assay,
  • Applicant discloses methods of performing assays.
  • a method of performing a clinical a ssay by a. clinical testing facility on a sample obtained from a subject comprising: Providing an order form listing a plurality of assays which may be ordered, wherem the cost of each assay of said plurality of assays is provided on said order form; Marking the order form to indicate the name and other identifying informat on of said subject; Marking the order form to identify one or more selected assays of the plurality of assays that are ordered for performance; Providing the marked order form to the clinical testing facility; Obtaining a sample from said subject during a single sample acquisition session; and Performing said one or more selected assays.
  • the sample is a small-volume biological sample having a volume of less than about 250 microliters i u i . s.
  • the sample is a blood sample or a urine sample.
  • the one or more selected assays comprise at least two linked assays, wherein said linked assays comprise an initial assay and at least one subsequent assay, wherein the results of said initial assay affect the decision whether or not to perform said at least one subsequent assay, or affect the decision whether or not to report results of said at least one subsequent assay.
  • the sample is provided in a sampl e container for performance of said assays by the clinical testing facility using an automated sample analysis device,
  • Further embodiments of these methods comprise performing a plurality of assays comprising an initial assay and a subsequent assay, wherem the sample is provided in a sample container that is disposed on a cartridge during placement of the sample within an automated sample analysis device, further comprising dividing said sample into portions, said portions comprising a first portion and at least a second portion, and wherein said second portion is retained in said sample container disposed on the cartridge within the device for use in the subsequent assay.
  • the sample is obtained from said subject during a single sample acquisition session.
  • the sample is a blood sample or a urine sample.
  • the cartridge contains all consumables used in the performance of the initial assay and of the at least one subsequent assay.
  • Applicant discloses further methods of ordering assays.
  • Applicant discloses a method of ordering at least an initial clinical assay and a subsequent clinical assay for performance by a clinical testing facility on a sample obtained from a subject, wherein said initial assay and said subsequent assay each comprise detecting, identifying, or measuring an analyte or characteristic of said sample, the method comprising: Providing an order form listing a plurality of assays which may be ordered, wherein the cost of each assay of said plurality of assays is provided on said order form, and wherein said plurality of assays includes linked assays, wherein said linked assays comprise an initial assay and at least one subsequent assay, wherein the performance of said at least one subsequent assay is contingent upon the results of said initial assay : Marking the order form to indicate the name and other identifying informat on of said subject; Marking the order form to indicate linked assays selected for performance, wherein said initial assay of said linked assays is to be performed on the sample or
  • Applicant discloses further methods of ordering assays.
  • Applicant discloses a method of ordering at least two clinical assays to be performed by a clinical testing facility on a sample obtained from a subject, said at least two clinical assays comprising finked assays, wherein linked assays comprise an initial assay and at least one subsequent assay, wherein said linked assays each comprise detecting, identifying, or measuring an analyte or characteristic of said sample, the method comprising: Providing an order form listing a plurality of assays which may be ordered, wherein the cost of each assay of said plurality of assays is provided on said order form, and wherein said plurality of assays includes linked assays, wherein said finked assays comprise an initial assay and at least one subsequent assay; Marking the order form to indicate the name and other identifying information of said subject; Marking the order form to indicate linked assays that have been selected for performance; wherein said initial assay and said at least one subsequent assay are to be performed
  • Methods of ordering may further comprise methods as disclosed above, wherein the at least one subsequent assay comprises an assay for detecting, identifying, or measuring: a) the same analyte or characteristic as the initial assay, and wherein the at least one subsequent assay is more sensitive for the detection or measurement of the analyte or characteristic than the initial assay, or b) a different analyte or characteristic than the analyte or characteristic detected, identified, or measured by the initial assay.
  • the methods may further comprise methods wherein the initial assay comprises an assay of an assay type selected from the group of assay types consisting of antibody-based assays, nucleic acid assays, general chemistry assays, and cytometric assays.
  • the methods may further comprise methods wherein the at least one subsequent assay comprises a different type of assay than the initial assay.
  • the methods may further comprise methods wherein said cytometric assay comprises the detection, identification, or measurement of a morphological characteristic of a cell.
  • the methods may further comprise methods wherein said cytometric assay comprises the detection, identification, or measurement of a morphological characteristic of a cell.
  • the methods may further comprise methods wherein the initial assay comprises the detection, identification, or measurement of an analyte, and the subsequent assay comprises a cytometric assay for the detection, identification, or measurement of a morphological characteristic of a cell
  • the methods may further comprise methods wherein the first portion of the sample is diluted prior to, or during, the performance of the initial assay, or wherein the second portion of the sample is diluted prior to, or during, the performance of the at least one subsequent assay, or both.
  • the sample is a sniall-voiume biological sample having a volume of less than about 250 microliters (uL).
  • the sample is provided in a sample container for performance of said assays by the clinical testing facility using an automated sample analysis device, the methods further comprising: dividing the sample into portions prior to the performance of said initial assay, wherein said portions comprise a first portion and at least a second portion.
  • the sample in said sample container is disposed on a cartridge during placement of the sample within said automated sample analysis device, and wherein said second portion is retained on said cartridge within the automated sample analysis device for use in the at least one subsequent assay.
  • the second portion is retained in said sample container disposed on the cartridge within the automated sample analysis device for use in the performance of the at least one subsequent assay.
  • the second portion is retained in a second container th t is disposed on the cartridge within the automated sample analysis device.
  • the second container is a dedicated container designated for holding the second portion on the cartridge prior to the performance of the at least one subsequent assay.
  • the second container is a dedicated container designated to contain the second portion during the performance of the at least one subsequent assay .
  • the performance of the subsequent assay is begun within a short amount of time after placing the sample within the automated sample analysis device, wherein the short amount of time is about two hours or less.
  • the automatic performance of the subsequent assay is completed within a short amount of time after placing the sample within the automated sample analysis device, wherein said short amount of time is about one hour or less.
  • the sample is provided on a cartridge, and wherein said cartridge contains all consumables used in the performance of the initial assay and of the subsequent assay, in embodiments of such methods, the first portion of the sample is diluted by at least about 10- fold prior to, or during, the performance of the initial assay, or wherein the second portion of the sample is diluted by at least about 10-foid prior to, or during, the performance of the subsequent assay, or both, wherein, following dilution, the volume of said diluted portion is less than about 200 microliters (uL).
  • the first portion of the sample is diluted by at least about 100-fold prior to, or during, the performance of the initial assay, or wherein the second portion of the small- volume sample is diluted by at least about 100-fold prior to, or during, the performance of the subsequent assay, or both, wherein, following dilution, the volume of said diluted portion is less than about 200 microliters (uL).
  • the initial assay comprises an assay of an assay type selected from the group of assay types consisting of antibody-based assays, nucleic acid assays, general chemistry assays, and cytometric assays.
  • the subsequent assay comprises a different type of assay than the initial assay.
  • cytometric assays comprise the detection, identification, or measurement of a morphological characteristic of a ceil
  • the subsequent assay comprises an assay for detecting, identifying, or measuring: a) the same analyte or characteristic as the initial assay, wherein the subsequent assay is more sensitive for the detection, identification, or measurement of the analyte or characteristic than the initial assay, or b) a different analyte or characteristic than the analyte or characteristic detected, identified, or measured by the initial assay.
  • the sample comprises a volume of less than about 100 microliters (uL).
  • the sample comprises a sample selected from the group of biological sample types consisting of blood, serum, plasma, a throat swab, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid, tears, stool, mucus, sweat, earwax, oil, a glandular secretion, semen, vaginal fluid, interstitial fluids derived from tumorous tissue, ocular fluids, breath, hair, finger nails, skin, biopsy tissue, placental fluid, amniotic fluid, cord blood, lymphatic fluids, cavity fluids, sputum, pus, microbiota, meconium, breast milk, and other secretions or excretions.
  • biological sample types consisting of blood, serum, plasma, a throat swab, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid, tears, stool, mucus, sweat,
  • the results of the initial assay of the sample are to be reported.
  • the results of the mitial assay are to be reported prior to the performance of the subsequent assay.
  • the results of the at least one subsequent assay are to be reported.
  • the results of the initial assay and of the at least one subsequent assay are to be reported together.
  • the linked assays comprising an initial assay and at least one subsequent assay are selected from the group of linked assays consisting of: An initial assay comprising a complete blood count with automated differential, and a subsequent assay comprising manual differential and smear review; An initial assay comprising a complete blood count with automated differential, and a subsequent assay comprising an assay for anemia; An initial assay comprising an acute hepatitis panel, and a subsequent assay comprising a quantitative nucleic acid assay for Hepatitis C; An initial assay comprising one or more of celiac panel assays selected from IgA and HLA-DQ typing, and a subsequent assay comprising one or more of celiac reflex panel assays selected from tTG, DGP, EMA, and EMA titer; An initial assay comprising a Hepatitis C antibody assay, and a subsequent assay comprising a quantitative nucleic acid assay for Hepati
  • a method of performing assays comprises a method of performing, by a clinical testing facility, at least an initial clinical assay and a subsequent clinical assay on a sample obtained from a subject, wherein said initial assay and said at least one subsequent assay each comprise detecting, identifying, or measuring an analyte or characteristic of said sample, the method comprising: Providing an order form listing a plurality of assays which may be ordered, wherein the cost of each assay of said plurality of assays is provided on said order form, and wherein said plurality of assays includes linked assays, wherein said linked assays comprise an initial assay and at least one subsequent assay, wherein the performance of said at least one subsequent assay is contingent upon the results of said initial assay; Marking the order form to indicate the name and other identifying information of said subject; Marking the order form to indicate linked assays selected for performance; Providing the marked order form to the clinical testing facility
  • a method of performing assays comprises a method of performing, by a clinical testing facility, at least two linked assays on a sample obtained from a subject, said linked assays comprising an initial assay and at least one subsequent assay, each of said linked assays comprising detecting, identifying, or measuring an analyte or characteristic of said sample, the method comprising: Providing an order form listing a plurality of assay s which may be ordered, wherein the cost of each assay of said plurality of assays is provided on said order form, and wherein said plurality of assays includes linked assays, wherein said linked assays comprise an initial assay and at least one subsequent assay; Marking the order form to indicate the name and other identifying information of said subject; Marking the order form to indicate linked assays that have been selected for performance; Providing the marked order form to the clinical testing facility; Obtaining a sample from said subject at a single sample
  • the at least one subsequent assay comprises an assay for detecting, identifying, or measuring: a) the same anaiyte or characteristic as the initial assay, and wherein the at least one subsequent assay is more sensitive for the detection or measurement of the anaiyte or characteristic than the initial assay, or b) a different anaiyte or characteristic than the anaiyte or characteristic detected, identified, or measured by the initial assay.
  • the initial assay comprises an assay of an assay type selected from the group of assay types consisting of antibody-based assays, nucleic acid assays, general chemistry assays, and cytometric assays.
  • the at least one subsequent assay comprises a different type of assay than the initial assay.
  • the cytometric assay comprises the detection, identification, or measurement of a morphological characteristic of a cell.
  • the initial assay comprises the detection, identification, or measurement of an anaiyte
  • the subsequent assay comprises a cytometric assay for the detection, identification, or measurement of a morphological characteristic of a cell.
  • the first portion of the sample is diluted prior to, or during, the perfonnance of the initial assay, or wherein the second portion of the sample is diluted prior to, or during, the perfonnance of the at least one subsequent assay, or both.
  • the sample is a small-volume biological sample having a volume of less than about 250 microliters ( ⁇ ,).
  • the sample is provided in a sample container for performance of said assays by the clinical testing facility using an automated sample analysis device, further comprising: Dividing the sample into portions prior to the performance of said initial assay, said portions comprising a first portion and at least a second portion.
  • the sample in said sample container is disposed on a cartridge during placement of the sample within said automated sample analysis device, and wherein said second portion is retained on said cartridge within the automated sample analysis device for use in the at least one subsequent assay.
  • said second portion is retained in said sample container disposed on the cartridge within the automated sample analysis device for use in the performance of the at least one subsequent assay.
  • said second portion is retained in a second container that is disposed on the cartridge within the automated sample analysis device.
  • said second container is a dedicated container designated for holding the second portion on the cartridge prior to the performance of the at least one subsequent assay.
  • the second container is a dedicated container designated to contain the second portion during the performance of the at least one subsequent assay.
  • the performance of the subsequent assay is begun within a short amount of time after placing the sample within the automated sample analysis device, wherein the short amount of time is about two hours or less.
  • the automatic performance of the subsequent assay is completed within a short amount of time after placing the sample within the automated sample analysis device, wherein said short amount of time is about one hour or less.
  • the sample is provided on a cartridge, and wherein said cartridge contains all consumabl es used in the performance of the initial assay and of the subsequent assay.
  • the first portion of the sample is diluted by at least about 10- fold prior to, or during, the performance of the initial assay, or wherein the second portion of the sample is diluted by at least about 10-fold prior to, or during, the performance of the subsequent assay, or both, wherein, following dilution, the volume of said diluted portion is less than about 200 microliters (uL).
  • the first portion of the sample is diluted by at least about 100-fold prior to, or during, the performance of the initial assay, or wherein the second portion of the small-volume sample is diluted by at least about 100-fold prior to, or during, the performance of the subsequent assay, or both, wherein, following dilution, the volume of said diluted portion is less than about 200 microliters (uL).
  • the initial assay comprises an assay of an assay type selected from the group of assay types consisting of antibody -based assays, nucleic acid assays, general chemistry assays, and cytometric assays.
  • the subsequent assay comprises a different type of assay than the initial assay.
  • the cytometric assays comprise the detection, identification, or measurement of a morphological characteristic of a cell.
  • the subsequent assay comprises an assay for detecting, identifying, or measuring: a) the same analyte or characteristic as the initial assay, wherein the subsequent assay is more sensitive for the detection, identi ication, or measurement of th analyte or characteristic than the initial assay, or b) a different analyte or characteristic than the analyte or characteristic detected, identified, or measured by the initial assay.
  • the sample comprises a volume of less than about 100 microliters (uL).
  • the sample comprises a sample selected from the group of biological sample types consisting of blood, serum, plasma, a throat swab, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, cerebrospinal fluid, tears, stool, mucus, sweat, earwax, oil, a glandular secretion, semen, vaginal fluid, interstitial fluids derived from tumorous tissue, ocular fluids, breath, hair, finge nails, skin, biopsy tissue, placental fluid, amniotic fluid, cord blood, lymphatic fluids, cavity fluids, sputum, pus, microbiota, meconium, breast milk, and other secretions or excretions.
  • the methods of performing assays further comprise:
  • reporting the results of the initial assay of the sample In embodiments, the results of the initial assay are reported prior to the performance of the subsequent assay. In embodiments, the methods comprise reporting the results of the subsequent assay. In embodiments, the results of the initial assay and of the subsequent assay are reported together.
  • the linked assays comprising an initial assay and at least one subsequent assay are selected from the group of linked assays consisting of: An initial assay comprising a complete blood count with automated differential, and a subsequent assay comprising manual differential and smear review; An initial assay comprising a complete blood count with automated differential, and subsequent assay comprising an assay for anemia; An initial assay comprising an acute hepatitis panel, and a subsequent assay comprising a quantitative nucleic acid assay for Hepatitis C; An initial assay comprising one or more of celiac panel assays selected from IgA and HLA-DQ typing, and a subsequent assay comprising one or more of celiac reflex panel assays selected from t ' TG, DGP, EMA, and EMA titer; An initial assay comprising a
  • Hepatitis C antibody assay, and a subsequent assay comprising a quantitative nucleic acid assay for Hepatitis C An initial assay comprising an HIV antibody screening assay, and a subsequent assay comprising a confirmatory assay for HIV; An initial assay comprising an HIV antibody screening assay, and a subsequent assay comprising an assay for differentiating between HIV- 1 antibodies and HIV-2 antibodies; An initial assay comprising an assay for Treponema pallidum IgG, and a subsequent assay comprising a rapid plasma reagin (RPR) assay; An initial assay comprising a rapid plasma reagin (RPR) assay, and a subsequent assay comprising an assay for an assay for antibodies to Treponema pallidum; An initial assay comprising a thyroid stimulating hormone (TSH) assay, and a subsequent assay comprising comprising one or more of an assay for rri-iodomyronine (T3), thyroxine (T4),
  • the sample is obtained in a single sample acquisition session, and wherein said sample comprises a volume sufficient for the perfoiTnance of an initial assay and a subsequent assay.
  • the sample is a small- volume sample having a volume of less than about 250 microliters (uL).
  • the sample is a small-volume sample having a volume of less than about 100 microliters ( ⁇ ).
  • the sample is a small- volume blood sample or a small- volume urine sample.
  • the sample is a blood sample obtained by fmgerstick.
  • Applicant further provides order forms (also termed “order sheet”, “lab order form”, “lab order sheet”, “assay form”, “assay sheet”, “assay order form”, “assay order sheet”, “test form”, “test sheet”, “test order form”, “test order sheet”, “medical order form”, “medical order sheet”, “medical test sheet”, “medical test sheet”, and plurals and similar and related terms) configured for ordering clinical diagnostic assays for testing biological samples obtained from human subjects.
  • order forms for ordering a clinical assay having features as disclosed herein are shown in Figs. 5A, 5B, and 6.
  • the order forms for ordering a clinical assay provide lists of clinical assays with prices for each clinical assay.
  • the list of clinical assays identifies each assay by at least one of: an assay name; an identifying acronym; an identifying number; an identifying alphanumeric designation; or combinations thereof.
  • the order forms provide lists of panels of clinical assays with prices for each panel.
  • the order forms provide lists of clinical assays with prices for each clinical assay, where the clinical assays that are listed include initial assays and corresponding subsequent assays, and corresponding prices, with explicit indications of the connection between such initial assays and corresponding subsequent assays.
  • the corresponding prices listed include prices for the subsequent assays and for the initial assays: in embodiments, a combined price may be listed for an initial assay and a subsequent assay.
  • prices are listed for panels of assays, which may include initial assays and subsequent assays.
  • the order forms pro vide lists of clinical assay s with prices for each clinical assay, where the clinical assays that are listed include initial assays and corresponding subsequent assays, and prices for the initial and corresponding assays, with explicit indications of the connection between such initial assay s and corresponding subsequent assays.
  • the listed prices include prices for the subsequent assays and for the initial assays; in embodiments, a combined price may be listed for an initial assay and a subsequent assay.
  • the order forms include lists of assays performed together in panels; in embodiments, the order forms include explanations of the assays to be performed, including explanations of the panels or of the assays performed together in panels.
  • some or all of the more commonly performed assays may be listed near to each other, or together, on the order forms.
  • some or all of the assays listed on the order forms may be listed near to, or grouped together with, other assays which may commonly be ordered for patients exhibiting a common symptom or common symptoms.
  • some or all of the assays listed on the order forms may be listed near to, or grouped together with, other assays which may commonly be ordered for patients sharing common risk factors.
  • some or all of the assays listed on the order forms may be listed near to, or grouped together with, other assays which may commonly be ordered for patients sharing common age, sex, activity, or other characteristic. In embodiments, some or all of the assays listed on the order forms may be listed alphabetically.
  • the order forms include check boxes for selecting and indicating assays to be performed (including reflex assays which may be performed, continent upon the results of initial assays); lines, boxes, or other locations for providing patient identification and contact information: lines, boxes, or other locations for providing clinician information; lines, boxes, or other locations for providing payer information (e.g., patient payment information, insurance company information, or other information regarding payment); lines, boxes, or other locations for providing clinic information; lines, boxes, or other locations for providing reimbursement or other code information (e.g., lCD-9, ICD-9- CM, ICD- 10, lCD-10-CM, TCPM, DSM, CPT, or other code for classifying a disease, treatment, test, or for insurance reimbursement; etc.).
  • the samples may include blood samples, urine samples, and other biological samples.
  • Applicant discloses a method of providing a clinical assay performed on a sample obtained from a subject, comprising: Providing an order form listing clinical assays and prices for each of said climcai assay s; Selecting one or more clinical assays from the order form; Obtaining a sample from the subject; Performing the selected clinical assay or assays; and Collecting payment for the selected clinical assay or assays, where the payment is determined by the price for each clinical assay listed on the order form.
  • the order form may comprise any order form discussed or shown herein.
  • the clinical assay or assays may be performed according to any method disclosed herein.
  • the clinical assays listed on the order form are listed in panels, and the order forms comprise prices for each panel.
  • the prices for each panel comprise capped prices for at least one panel , wherein a capped price is a maximum price for any combination of individual clinical assays of a panel.
  • the payment is collected, at least in part, from the subject, or from an insurance company, or from a government agency, or from the subject's employer, or any combination thereof.
  • Applicant discloses a further method of providing a clinical assay performed on a sample obtained from a subject, comprising: Providing an order form listing climcai assays, prices for each of said clinical assays, and CPT codes for each of said clinical assays; Selecting one or more clinical assays from the order form; Obtaining a sample from said subject; Performing the selected clinical assays; and Collecting payment for the selected clinical assay or assays, where the payment is determined by the price for each clinical assay listed on the order form.
  • the order form may comprise any order form described or shown herein.
  • the clinical assay may be performed according to a method as disclosed herein.
  • the clinical assays listed on the order form comprise clinical assays listed in panels, and the order forms comprise prices and a CPT code or CPT codes for each panel.
  • the prices for each panel comprise capped prices for at least one panel, wherein a capped price is a maximum price for any combination of individual clinical assays of a panel.
  • the payment is collected, at least in part, from the subject, or from an insurance company, or from a government agency, or from the subject's employer, or any combination thereof.
  • Applicant further discloses a method of doing business, comprising: Providing an order form listing clinical assays and prices for each of said clinical assays; Selecting one or more clinical assays from the order form; Obtaining a sample from a subject; Performing the selected clinical assay or assays; and Collecting payment for the selected clinical assay or assays, where the payment is determined by the price for each clmical assay listed on the order form.
  • the order form may comprise any order form discussed or shown herein.
  • the clinical assays may be performed according to any method disclosed herein.
  • the clinical assays listed on the order form comprise clinical assays listed in panels, and the order forms comprise prices for each panel.
  • the prices for each panel comprise capped prices for at least one panel, wherein a capped price is a maximum price for any combination of individual clinical assays of a panel.
  • the payment is collected, at least in part, from the subject, or from an insurance company, or from a government agency, or the subject's employer, or any combination thereof.
  • Applicant further discloses a method of doing business, comprising: Providing an order form listing clinical assays, prices for each of said clinical assays, and CPT codes for each of said clinical assays: Selecting one or more clinical assays from the order form;
  • the order form may comprise any order form discussed or shown herein.
  • the clinical assays may be performed according to any method disclosed herein.
  • the clinical assays listed on the order form comprise clinical assays listed in panels, and the order form comprises prices and CPT codes for each panel.
  • prices for each panel comprise capped prices for at least one panel, wherein a capped price is a maximum price for any combination of individual clinical assays of a panel.
  • the payment is collected, at least in part, from the subject, or from an insurance company, or from a government agency, or from the subject's employer, or any combination thereof.
  • Example 1 - HIV Assays for the detection of human immunodeficiency virus (HIV) in a biological sample typically report negative results, i.e., no HIV is detected in the sample. However, since detection of the vims in a biological sample is of such importance, and may have such dire consequences for the subject from whom the sample is obtained, Applicant discloses methods for performing confirmatory follow-up testing on the basis of positive results to an initial assay. In embodiments, such confirmatory follow-up testing may be performed prior to reporting positive assay results to a health-care provider or to a subject. In embodiments, such confirmatory follow-up testing may be performed concurrent with, or following, the reporting of positive assay results to a health-care provider or to a subject.
  • confirmatory follow-up testing may be performed prior to reporting positive assay results to a health-care provider or to a subject.
  • an anti-HIV antibody screening test may be performed on a biological sample obtained from a subject; for example, an anti-HIV antibody screening test may be performed on a sample of blood obtained from a subject.
  • an anti- HIV antibody screening test may be performed on a sample of a bodily fluid obtained from a subject other than a blood sample; such a sample may be, for example, a sample of urine, sputum, semen, tears, interstitial fluid, a sample obtained from a nasal swab, a sample obtained from a throat swab, a sample obtained from a vaginal swab, or other sample.
  • a sufficient amount of a biological sample is obtained so as to allow the automatic reflex testing of the sample, or of a portion of the sample, without need to obtain a further sample in order to perform a subsequent assay.
  • only the amount of a biological sample needed for an initial assay or assays is obtained; in such an embodiment, a further biological sample must later be obtained if needed in order to perform a subsequent assay contingent on the results of an initial assay.
  • an antibody-based assay may comprise contacting a sample with a substrate to which antibodies specific for a target antigen are bound.
  • the sample may be mixed, or diluted, with a reagent containing a known amount of labeled conjugate, where the labeled conjugate binds the bound antibodies.
  • the labeled conjugate may be a conjugate comprising the target analyte covalentiy linked with a detectable label.
  • the mixture of sample and reagent may be then added to a chamber containing the substrate to which the antibodies are bound, and left to incubate for a sufficient time for target analyte, and analyte-conjugate, to bind to the substrate.
  • the chamber may be washed with a washing solution, in order to wash out any remaining unbound analyte and unbound analyte- conjugate.
  • the amount of labeled conjugate bound to the substrate may be determined, and the amount of target analyte in the sample determined.
  • Such a determination may be made, e.g., by comparison with the amount of labeled conjugate bound in the absence of any target, and optionally by comparison with the amount of labeled conjugate bound in the presence of one or more known amounts of, or otherwise by comparison with control values, such as with a control curve.
  • a reagent allowing the detection of the label may be added to the chamber (e.g., where the label is an
  • the reagent allo wing the detection of the label may comprise the required substrate).
  • the anti-HIV antibody screening test may comprise an anti- HIV- l antibody screening test. In embodiments, the anti-HIV antibody screening test may comprise an anii-HiV-2 antibody screening test. In embodiments, the anti-HIV antibody screening test may comprise an anti-HIV- 1 and an anti-Ill V-2 antibody screening test.
  • the possible results of such an assay may comprise a result reporting that the sample is negati ve for the presence of HIV- 1 , and may comprise a result reporiing that the sample is positive for the presence of HIV-1 ; in such embodiments, a normal result comprises a result reporting that the sample is negative for the presence of HIV- 1.
  • the possible results of such an assay may comprise a resuit reporting that the sample is negative for the presence of HI V-2, and may comprise a result reporting that the sample is positive for the presence of HI V-2; in such embodiments, a normal resuit comprises a result reporting that the sample is negative for the presence of HIV-2.
  • the result of such a test comprises a result that the sample is negative for the presence of HIV- 1 , or negative for the presence of HIV-2
  • no further HIV test is automatically performed.
  • the HIV 7 test comprises testing for the presence of both HIV- 1 and HIV-2
  • the resuit of such a test comprises a resuit that the sample is negative for the presence of HIV- 1 and is negative for the presence of HIV-2
  • no further HIV test is automatically performed.
  • a further HIV test is automatically performed.
  • a further HIV test may be performed on the same sample, or on an additional sample.
  • the additional sample may be, or may be obtained from, a sample that was originally obtained from the subject; or, in embodiments, a further sample may be obtained from the subject and used for the further HTV test.
  • the further HIV test may comprise a nucleic acid assay.
  • the further HIV test may comprise a Western Blot HIV test.
  • the possible results of such a further HIV test may comprise a result reporting that (he sample is negative for the presence of HIV- 1 , or is negat ive for the presence of HIV-2, or is negative for the presence of both HIV-1 and HIV-2,
  • a normal result comprises a result reporting that the sample is negative for the presence of HIV
  • the result of a further HIV test comprises a result that the sample is negative for the presence of HIV-1 , or negative for the presence of HIV-2
  • no further HIV test is automatically performed.
  • results may be reported to a health-care provider, or to a subject, if appropriate, as indicating that the biological sample obtained from the subject is free of HIV.
  • results of such a further HIV test may comprise a result reporting that the sample is positive for the presence of HIV-1, or is positive for the presence of HIV-2, or is positive for the presence of both HIV-1 and HIV-2.
  • results of a further HIV test comprises a result that the sample is positive for the presence of HIV- 1 , or positive for the presence of HIV-2
  • a result indicates that HIV is present in the biological sample obtained from the subject.
  • results indicate that the subject may suffer from HI V.
  • results may be reported to a health-care provider, or to a subject, if appropriate, as indicating that the biological sample obtained from the subject contains HIV.
  • an initial assay may be an assay for the determination of the white blood cell count of a sample of blood obtamed from a subject.
  • white blood cell count may be obtained as part of a complete blood cell count.
  • a white blood cell count assay may determine ihai the white blood cell count of blood obtained from a subject falls outside a normal range, e.g., may be below about 2000 cells per microliter (fiL). Where an initial white blood cell count assay result determines that the white biood cell count of a blood sample is outside of a normal range, a reflex blood test may be required. Accordingly, Applicant discloses methods for performing confirmatory follow-up testing on the basis of white blood cell count results that fall outside of a pre-determined range, in embodiments, such confirmatory follow-up testing may be performed prior to reporting the initial white biood cell count results to a health-care provider or to a subject,
  • a normal range for an adult male subject may be between 3200 white blood eells. uL and 10,600 white blood
  • a reflex blood test may be performed.
  • a reflex blood test may comprise cytometric examination of the blood of the subject, such as a cytometric examination of white blood cells in a sample of blood from the subject.
  • the sample of blood examined in the reflex assay may be a portion of the sample of blood examined in the initial test, e.g., where a portion of the sample is retained for further testing.
  • the sample of blood examined in the reflex assay may be a sample of blood obtained at the same time as the sample of blood examined in the initial test, e.g., where a second blood sample is obtained and retained for further testing.
  • a subsequent sample of blood may be obtained from the subject for reflex testing at a time after the time of obtaining the initial sample of blood.
  • the subsequent sample of blood may be obtained following determination of the results of the initial assay.
  • an automatic cytometric assay may be used to identify, quantify, and classify white blood cells in a sample.
  • Blood samples may be pre-treated so as to avoid interference by red blood cells and platelets, e.g., by causing swelling and lysis of red blood ceils and platelets, allowing white blood ceils to settle and attach to a substrate, or by other means.
  • White blood cells may be contacted with one or more labels specific for cell markers, which are thus useful for identifying and classifying the white blood cells.
  • labels may include labels, such as fluorescent labels, to ease the detection of the labels and of cells labeled thereby.
  • Assays and tests that may be contingent on the results of an initial test may include assays and tests that observe and describe cells in a biological sample, including assays and tests that identify cells, that determine the numbers of cells of one or more populations of ceils, or that determine whether or not abnormal cells are present, or whether or not abnormal numbers of a cell type or cell types are present. Such tests and assays may utilize cytometry.
  • Cytometry may include preparing and analyzing two-dimensional images of cells in a biological sample, where the cells are labeled (e.g., with fluorescent,
  • chemifuminescent, enzymatic, or other labels and plated (e.g., allowed to settle on a substrate) and imaged by a camera.
  • the camera may include a lens, and may be attached to or used in conjunction with a microscope. Cells may be identified in the two-dimensional images by their attached labels (e.g., from light emitted by the labels),
  • the centrifuged capped vessel was loaded into a cartridge containing multiple fluidically isolated reagents, tips, and a cytometry cuvette.
  • the cartridge contained all the reagents required for the assay.
  • the cartridge was loaded into a. device equipped with at least a centrifuge, a pipette and a platform to load the cuvette.
  • the pipette in the device has a plurality of nozzles, some nozzles being of a different size than some other nozzles,
  • a nozzle on the pipette was lowered on a cuvette carrier tool causing it to engage a corresponding hole on the carrier tool. This tool was subsequently moved to the cartridge and lowered on the cytomeier cuvette. Pins on the tool were then able to engage corresponding holes on the cuvette and pick it up. The cuvette was transferred to a loading station elsewhere in the device.
  • N ext inside the device, a larger nozzle of the pipette was lowered into the cartridge to engage a pipette tip stored in the cartridge.
  • the pipette and tip together were then used to mix the cells and plasma in the capped vessel by positioning the pipette tip within the sample in ihe capped vessel and repeatedly aspirating material into and dispensing material from the tip.
  • 5 microliters of the mixed whole blood was aspirated to provide an aliquot for measurements of properties of the blood sample. This 5 microliter aliquot was used for measurements directed to the red blood cells and platelets in the sample. As discussed below r , a portion of the sample remaining after removal of this 5 microliter aliquot was used for measurements directed at white blood ceils in the sample.
  • the pipette then engaged a tip in the cartridge, aspirated 7 microliters ( ⁇ ) of the above mixture of and loaded the 7 ⁇ , into a channel within the cuvette placed on a platform with the carrier tool.
  • the pipette aspirated 10 ⁇ of mineral oil from a vessel in the cartridge, and placed a drop of mineral oil on both open ends of the loaded channel of the cuvette.
  • Mineral oil w as added to the ends of the open channel to prevent evaporation of liquid from the loaded cuvette channel .
  • the device-level sample handling apparatus engaged the cuvette carrier / cuvette combination, and transported the cuvette carrier / cuvette combination from the module containing the cartridge to the cytometry module of the device.
  • the device-level sample handling apparatus placed the cuvette carrier / cuvette combination on the microscopy stage of the cytometry module.
  • the time required for these operations, in addition to a 2 minute wait time allowed the swollen ceils to settle to the floor of the cuvette prior to imaging.
  • the stage was moved to pre-detennined location so that the optical system of the cytometer could view one end of the channel containing the sample.
  • the optical system relayed images of the sample acquired with darkfield illumination from a ringlight. These images coupled with actuation of the optical system on an axis perpendicular to the plane of the cuvette were used to find the plane of best focus. Once focused, the optical system was used to acquire fluorescence images of the sample at different wavelengths, commensurate with the fluorophores that were being used.
  • red blood cells that had been labeled with anti-CD235 conjugated to alexa fiuor 647
  • a red (630 nanometer (nm) wavelength) light source was used to excite the sample and wavelengths between 650nm and 700nm were used to image the sample.
  • a combination of a polychroic mirror and a bandpass emission filter was used to filter out unwanted wavelengths from the optical signal Since the cells had settled on the floor of the cuvette, images at a single plane of focus were sufficient to visualize all cells in the region.
  • Data from the images was processed by a controller associated with the automated assay device.
  • the image processing algorithms employed here utilized fluorescence images of cells to detect them using a combination of adaptive thresholding and edge detection. Based on local intensity and intensity gradients, regions of interest (Rol) were created around each cell. Using darkfield images, beads in the sample were also identified and Rols were created around the beads. All the Rols in each field of view were enumerated and their intensity in each image of that field of view were calculated.
  • the information output by the image processing algorithm consisted of shape or morphometric measurements and fluorescence and darkfield intensities for each Rol.
  • This information was analyzed using statistical methods to classify each object as either a red blood cell (positive for CD235a, but negative for CD41/CD61), a. platelet (positive for CD41/CD61 and negative CD235a) or a bead.
  • the shape descriptors such as perimeter, diameter and circularity were used to calculate the volume of each red blood cell and platelet. Since the beads were added at a known concentration, the average ratio of beads to cells over the whole channel was used to calculate cell concentration in terms of cells/microliter. Based on the steps performed for processing the sample, this concentration was corrected for dilution to arrive at concentration of cells in the original whole blood sample.
  • the following quantities were calculated from a sample: 1) number of red blood cells in the cuvette; 2) average v olume of red blood ceils in the cuvette; 3) red blood cell distribution width (RDW) of red blood cells in the cuvette; 4) number of platelets in the cuvette; and 5) average volume of platelets in the cuvette. Based on these calculations, the following was calculated for the original blood sample.
  • the remaining 75 ⁇ iL of sample was used to analyze the white blood cell population of the whole blood sample.
  • the remaining 75 iiL of whole blood had also been mixed by repeatedly aspirating and dispensing the sample within the same the vessel by the pipette.
  • Approximately 40 ⁇ , of the remaining 75 iiL of mixed whole blood was aspirated into a pipette tip, and transferred by the pipette to a centrifuge tube in the cartridge.
  • the centrifuge tube containing the blood sample was engaged by the pipette, and transferred to and deposited in a swinging bucket in a centrifuge within the module.
  • the centrifuge was spun to provide 1200 x g for 3 minutes, separating the blood into EDTA-containing plasma as the supernatant and packed cells in the pellet.
  • the centrifuge tube was removed from the centrifuge and returned to the cartridge.
  • the plasma supernatant was removed by the pipette and transferred to a separate reaction vessel in the cartridge.
  • 16 ,uL of resuspension buffer was aspirated by the pipette, and added to the cell pellet in the centrifuge tube.
  • the pipette then resuspended the cell pellet in the resuspension buffer by repeatedly aspirating and dispensing the mixture in the centrifuge tube.
  • the pipette aspirated 21 microliters of the resuspended whole blood and added it to another vessel containing 2 microliters of anti-CD 14-pacific blue and DRAQ5 ® , mixed, and incubated for 2 minutes. Twenty microliters of this mixture was then added to 80 microliters of a lysis buffer.
  • the lysis buffer is a solution of a gentle surfactant such a saponin in conjunction with a fixative such as paraformaldehyde.
  • the detergent causes a large number of holes to be formed in the membranes of cells. Red blood ceils, due to their unique membrane properties, are particularly susceptible to this hole formation and lyse completely, their contents leaking out into the liquid around.
  • Presence of the fixative prevents unintentional lysis of the white blood cells. Platelets also remain uniysed. The purpose of this step is to remo v e red blood cells from the mixture as they outnumber white blood cells by about 1000: 1. Platelets do not interfere with imaging and hence are irrelevant to this process.
  • the lysis buffer also contained 10 ⁇ non-fluorescent beads at a known concentration.
  • the vessel was spun again at 1200 x g for 3 minutes.
  • the supernatant was aspirated by a pipette tip, removing the red blood cell ghosts and other debris, and deposited into a waste area in the cartridge. Approximately 15 ⁇ , of liquid with packed white blood cells were present in the ceil pellet.
  • the pipette in order to determine a rough approximation of the number of white blood cells present in the cell pellet, the pipette first resuspended the white blood cells in the vessel and then aspirated the liquid, transferred it to spectrophotometer in the blade
  • the white blood cell suspension was illuminated with light at a wavelength of 632. nm, which is the excitation wavelength for alexa fiuor 647 dye and DRA.Q5 ® .
  • the light emitted by the cell suspension was filtered by a 650 nm long pass filter and measured in the spectrophotometer. This measurement was correiaied with previously generated calibration curve to estimate a rough concentration of white blood cells in the cell suspension.
  • cell concentrations ranged from about 1000 cells per iiL to about 100,000 cells per ⁇ iL. This estimate was used to calculate an appropriate dilution factor to ensure that the concentration of cells in the cuvette was constrained to within a two-fold range around a pre-defined target concentration. The purpose of this step was to ensure that cells are not present at too high or too low a density on the cuvette. If the cell density is too high, the accuracy of image processing algorithms is compromised, and if the cell density is too low, an insufficient number of cells are sampled.
  • the device-level sample handling apparatus engaged the cuvette carrier / cuvette combination, and transported the cuvette carrier / cuvette combination from the module containing the cartridge to the cytometry module of the device.
  • the de vice-level sample handling apparatus placed the cuvette carrier / cuvette combination on the microscopy stage of the cytometry module. After the cuvette carrier / cuvette was placed on the microscopy stage, the two channels of the cuvette containing white blood cells were imaged as described above for the RBC / platelet mixture.
  • Fig. 1A Darkiield images of the white blood cells were used to count the numbers of cells in a field (as shown in Fig. 1A).
  • Cell surface markers were used to determine the cell type of individual white blood cells in an image; for example, CD 14 marks monocytes; CD 123 marks basophils; CD 16 marks neutrophils; and CD45-AF647 were used to mark all leukocytes (Figs. I B- IE).
  • the nuclear stain DRAQ5 ® was used to mark cell nuclei, and so to differentiate nucleate cells (such as white blood cells) from mature red blood cells, which have no nucleus.
  • the image processing algorithms employed here utilized fluorescence images of cells to detect them using a combination of adaptive thresholding and edge detection. Based on local intensity and intensity gradients, boundaries of regions of interest (Rol) were created around each cell. Using darkiield images, beads in the sample were also identified and Rol boundaries were created around the beads. All the Rois in each field of view were enumerated and their intensity in each image of that field of view were calculated.
  • the information output by the image processing algorithm consisted of shape or morphometric measurements and fluorescence and darkiield intensities for each Rol. This information was analyzed using statistical methods to classify each object as a lymphocyte, monocyte, basophil, eosinophil, neutrophil or a bead.
  • Figs. 1 , 2, and 3 Examples of images and plots of results of such measurements are presented in Figs. 1 , 2, and 3.
  • Fig. 1 shows representative images of blood cells from a sample of whole blood; these images were taken using different imaging techniques and dyes.
  • the image shown in Fig. 1A was taken of cells from whole blood using dark-field illumination.
  • the image shown in Fig, IB was taken of cells from whole blood showing fluorescence from anti- CD 14 antibodies labeled with PAC Blue dye; the fluorescent cells are monocytes.
  • the image shown in Fig, 1 C was taken of cells from whole blood showing fluorescence from anti- CD 123 antibodies labeled with PECy5 dye; the fluorescent cells are basophils.
  • the image shown in Fig, ID was taken of cells from whole blood showing fluorescence from anti-CD 16 antibodies labeled with PE dye; the fluorescent cells are neutrophils.
  • the image shown in Fig. IE was taken of cells from whole blood showing fluorescence from anti-CD45 antibodies labeled with AF647 dye; ail leukocytes fluoresce under these conditions.
  • the image shown in Fig. IF was taken of cells from whole blood dyed with DRAQ5 ® to stain cell nuclei. ' T ' hus, leukocytes and platelets are stained and fluoresce under these conditions, but red blood cells (lacking nuclei) are not stained and do not fluoresce.
  • Fig. 2 shows a representative composite image of cell-types in whole blood from images acquired according to the methods disclosed herein. Images of a monocyte (labeled and seen in the upper left quadrant of the figure, with a reddish center surrounded by a blue-purple ring), a lymphocyte (labeled and seen in the center of the figure, with a bright red center surrounded by a dimmer red ring), an eosinophil (labeled and seen in the lower left quadrant of the figure, with a green center surrounded by a red border), and a neutrophil (labeled and seen in the lower right quadrant of the figure, with a green center surrounded by a yellow and green border) are shown in the figure.
  • a monocyte labeleled and seen in the upper left quadrant of the figure, with a reddish center surrounded by a blue-purple ring
  • a lymphocyte labeleled and seen in the center of the figure, with a bright red center surrounded by a dimmer
  • Fig, 3 shows plots of various cell types identified and quantified by the cytometric assays described in this example.
  • Fig. 3A shows a plot of spots (cells) by intensity of the marker F ' L- 17 versus intensity of the marker FL-9 to identify monocytes.
  • Fig. 3B shows a plot of spots (cells) by intensity of the marker FL- 19versus intensity of the marker FL- 15to identify basophils.
  • Fig. 3C shows a plot of spots (cells) by intensity of the marker FL- 15 versus intensity of the marker FL- 1 1 to identify lymphocytes.
  • Fig. 3D shows a plot of spots (cells) by intensity of the marker FL-15 versus intensity of the marker FL-9 to identify neutrophils and eosinophils.
  • the initial identification of monocytes (9.6%, as shown in Fig. 3A) is used to guide the subsequent identification of basophils (0.68%, as shown in Fig. 3B).
  • the identification of monocytes and basophils as shown in Figs. 3A and 3B is used to guide the subsequent identification of neutrophils and eosinophils (68% neutrophils, 3.2% eosinophils, of the WBCs shown in Fig. 3D).
  • lymphocytes are identified as shown in Fig. 3C (93% of the WBCs plotted in Fig. 3D, corresponding to 1 8% of the cells in the original sample).
  • the present methods correlate well with other methods. Counts of white blood cells, red blood cells, and platelets were made with samples of EDTA-anti coagulated whole blood. The white blood cells were further counted to determine the numbers of neutrophils, monocytes, and lymphocytes in the sample, in the measurements shown in Fig. 9, EDTA-anti coagulated whole blood samples were split into two, and one part of the samples were run on the system disclosed herein, using the methods disclosed herein. The other pari of the samples was run on an Abbott CELL-DYN Ruby System (Abbott
  • Fig. 4 shows plots demonstrating that cytometric methods as disclosed herein identify different cell types consistent with such identification by other methods.
  • WBCs white blood cells
  • RBCs red blood ceils
  • Fig. 4C platelets
  • Fig. 5 A shows one side of an order form for ordering blood tests that includes listings of test panels and of reflex tests according to the methods disclosed herein.
  • Fig. 5A includes lists many different tests which may be ordered for, and performed on samples of blood obtained from, a subject. These tests may be grouped together in panels, as shown in Fig. 5 A.
  • a metabolic panel may include (but need not be limited to) the following tests: BMP - Basic Metabolic Panel; CBC (complete blood count), no Diff; CBC with Automatic Differential; Celiac Panel; Celiac Panel Reflex; Chlaniydia/Gonorrhea; CMP - Comprehensive Metabolic Panel; Coagulation Panel with Reflex; Drug Panel;
  • Electrolytes Panel Epstein-Barr (EBV) Antibody Panel; Hepatic Function Panel; Hepatitis Panel, Acute; Lipid Panel; Obstetric Panel; Renal Function Panel; and STD Panel.
  • Diff and diff as used on an order form are abbreviations that refer to “Differentiation” assays; similarly, without regard to capitalization of any letters, the terms “auto diff” and “manual diff” as used on an order form are abbreviations that refer to “automatic differentiation” assays and “manual differentiation” assays.
  • a reflex test may be indicated; such a reflex test may include a Manual Diff & Smear Review test (e.g., a blood test in which both a manual count of different types of blood cells (such as different WBC types, and an inspection of a blood smear (e.g., a manual or visual inspection of a blood smear) is performed), an Anemia Test, or both.
  • a reflex test may be indicated; such a reflex test may include a celiac panel reflex test which provides more extensive or more sensitive testing, or both.
  • a reflex test may be indicated; such a reflex test may include a hepatitis C test, a quantitative RNA test (e.g., for hepatitis C RNA), or both.
  • Fig. 5A many other tests may be commonly ordered for subjects and performed on blood samples obtained from the subjects. Some of these tests include reflex tests where the results indicate that such further testing would be helpful. For example, as discussed above, and as indicated in Fig. 5A, a positive result of an HIV antibody screen (e.g., for HIV- 1 , H!V-2, or both) indicates, and may provoke, a reflex test for a confirmatory HIV test. Similarly, as discussed above, for example, as discussed above, and as indicated in Fig.
  • a positive result of a syphilis screen indicates, and may provoke, a reflex test for an antibody test (treponema pallidum "TP antibody") test that may serve as a confirmatory syphilis test.
  • An abnormal result of a thyroid stimulating hormone (TSH) test indicates, and may provoke, a reflex test for a confirmatory thyroid panel of tests confirmatory of, more sensitive for, thyroid abnormalities or thyroid hormone deficiencies than the initial TSH screen.
  • a basic metabolic panel includes tests for glucose, blood urea nitrogen (BUN), sodium (Na), potassium (K), chloride, (CI), carbon dioxide (C02), and calcium (Calcium Total, indicative of the level of total calcium (i.e., in all forms) in the blood sample).
  • a complete metabolic panel infudes tests for glucose, blood urea nitrogen (BUN), sodium (Na), potassium (K), chloride, (CI), carbon dioxide ( C02), and calcium (Calcium Total) as in the BMP, and further includes tests for albumin, alkaline phosphaiese (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (Bilirubin Total), and total protein (Protein Total).
  • an electrolyte panel includes tests for Na, K, CI, and C02,
  • a lipid panel includes tests for cholesterol (CtlOL), high density lipoprotein (HDL), low density lipoprotein ⁇ ! .[)! . ; ⁇ . and triglycerides (Trig).
  • CtlOL cholesterol
  • HDL high density lipoprotein
  • Trig triglycerides
  • a renal panel includes tests for glucose, BUN, creatinine, Na, K, CI, C02, Calcium Total, and albumin.
  • a complete blood count includes tests for hematocrit (HCT), hemoglobin (HGB), red blood cell count (RBC), white blood cell count (WBC), and platelet count.
  • HCT hematocrit
  • HGB hemoglobin
  • RBC red blood cell count
  • WBC white blood cell count
  • platelet count e.g., platelet count.
  • a blood test with differential e.g., a WBC count with differential provides a count for each of several different types of white blood cells.
  • a complete blood count (CBC) with reflex to anemia testing includes the CBC tests, and further includes (as reflex tests) tests for ferritin, folate, iron, total iron binding capacity (TIBC), vitamin B 12, and a smear review test (inspection of a blood smear).
  • a celiac panel includes tests for immunoglobulin A (IgA), and test for HLA-DQ typing (a serotype test). If indicated by the results of the celiac panel, further testing (reflex testing) including tests for tissue transglutaminadase antibody (tTG), deamidated gliadin peptide (DGP), and endomysial antibody (EMA) may be performed. Depending on the results of the EMA test, a reflex test for EMA titer may be performed.
  • a coagulation profile panel includes tests for platelet count (auto platelet count), prothrombin time and international normalized ratio test (PT/INR), partial thromboplastin time (PTT), and fibrinogen activity.
  • a drug panel includes iests for Amphetamines, Barbiturates, Benzodiazepines, Cocaine, Dofophine (methadone), Ecstasy, (MDMA), Marijuana (THC), Heroin (opiate screen), Methadone metabolite, Phencyclidine (PCP), Propoxyphene, and Tricyclic antidepressants.
  • an Epstein-Barr Antibody (Epstein-Barr Ah) test includes tests for immunoglobulin G (IgG) antibodies to Epstein-Barr early antigen (EA), IgG antibodies to Epstein-Barr nuclear antigen (NA), IgG antibodies to Epstein-Barr viral capsid protein (VCA), and immunoglobulin M (IgM) antibodies to Epstein-Barr VCA.
  • IgG immunoglobulin G antibodies to Epstein-Barr early antigen
  • NA Epstein-Barr nuclear antigen
  • VCA Epstein-Barr viral capsid protein
  • IgM immunoglobulin M antibodies to Epstein-Barr VCA
  • a hepatic function panel includes tests for albumin, ALP, ALT, AST, Bilirubin Direct, total bilirubin (Bilirubin Total), and total protein (Protein Total).
  • an acute hepatitis panel includes tests for hepatitis A virus (HAV), hepatitis B core antigen (HbcAn), hepatitis B surface antigen (HBsAg), and hepatitis C vims (HCV).
  • HAV hepatitis A virus
  • HbcAn hepatitis B core antigen
  • HBsAg hepatitis B surface antigen
  • HCV hepatitis C vims
  • an obstetric panel includes tests for ABO RhD, an antibody screen (Ab screen), a syphilis test (SYP), an HBsAg test, a rubella antibody test (RUB IgG), and a complete blood count with differential (CBC w/diff).
  • a sexually transmitted disease panel includes tests for gonorrhea and chlamydia (CT/'GC), hepatitis B surface antigen (HbsAg), hepatitis C virus (HCV), human immunodeficiency vims I and 2 (HIV 1/2), herpes simplex vims i and 2 (HSV 1/2), and Syphilis rapid plasma regain test (RPR).
  • CT/'GC gonorrhea and chlamydia
  • HbsAg hepatitis B surface antigen
  • HCV hepatitis C virus
  • HCV 1/2 human immunodeficiency vims I and 2
  • HSV 1/2 herpes simplex vims i and 2
  • RPR Syphilis rapid plasma regain test
  • a Triage Panel includes tests for BMP, amylase, PT/INR, prothrombin time (PIT), a CBC with auto differential and with reflex to manual differential and smear reviews of the blood sample.
  • a Thyroid stimulating hormone with reflex to Thyroid Panel includes tests for free tri-iodothyronine (free T3), free thyroxine (free T4), and a thyroid uptake test.
  • tests offered may be listed alphabetically (e.g., tests listed under the headings "Common Tests A-G", Common Tests H-S", and “Common Tests T-Z)"), listed according to type of sample (e.g., tests listed under "Stool” and “Urine”), grouped under the heading "Common Panels", or other groups.
  • a panel is a group of tests performed together; for example, as discusssed above, a basic metabolic panel (BMP) includes tests for glucose, blood urea nitrogen (BUN), sodium (Na), potassium (K), chloride, (CI), carbon dioxide (C02), and calcium (Calcium Total, indicative of the level of total calcium (i.e., in all forms) in the blood sample).
  • BMP basic metabolic panel
  • a lab order form as disclosed herein may include fines, spaces, boxes, or other locations for entering patient information, clinician information, insurance information, billing codes, and may include locations for particulars regarding testing (e.g., whether or not the tests are for a fasting patient; whether or not the test is urgent, or is routine; whether or not the test is to be performed repeatedly, and if so, at what frequency; and so forth).
  • Clinician information may include clinician name, NPI number, address, and other clinician information.
  • a lab order form may provide a location for a signature (e.g., for the signature of the ordering clinician).
  • a lab order form as illustrated in Fig. 5A may include a location to fill in a date after which the test order is no longer valid (e.g., "valid until" location).
  • test methodologies are embodied in the lab requisition form, in such an example, the design of the lab requisition form includes but is not limited to having prices on it and the way the tests are laid out on the form which is in part an algorithm that says if a subject has a certain test and has certain results, one should run certain other tesis,
  • the lab order form embodies algorithm-based testing across different assay and/or signal detection methodologies, such that many unique features incorporated into the layout of the lab requisition form.
  • one feature is the option of selecting whether all of the tests should be done by a micro-sample and by traditional phlebotomy.
  • some embodiments may designate which tests are done by micro-sample (e.g., a small-volume sample having a volume of about 250 or less) and which are done by other technique(s).
  • some may default to micro-sampling for most tests and only traditional phlebotomy using venipuncture for any remaining test(s) that require it due to technical or other limitations.
  • Another feature is an option to designate frequency (routine, urgent, standing order, and others) of testing.
  • a still further feature is a cash pay option, wherein a doctor or other lab reqisition ordering medical professional may opt to have the patient perform the test under cash pay, which essentially makes the test anonymous to health insurers. This may be of particular interest to patients who may have a certain lifestyle, condition, or have certain health risks where the subject may desire to test on a certain frequency (every quarter, every month, annually, or some other frequency, including just once) that if such testing and/or testing frequency is known to health insurers, may have a negative impact on their insurance coverage.
  • a still further feature is that prices are also shown for a substantial number, or optionally, all of the tests on the form.
  • RNA or other nucleic acid amplification-based screen Normally, when one does reflex testing, one may use an antibody screen and then reflex to an RNA or other nucleic acid amplification-based screen.
  • the antibody test describes mainly the immune response but the quantitative nucleic acid test pro vides information and measurements regarding viral load and can more definitively show whether a subject has the disease or not.
  • an algorithm-based lab order form one may test for CBC and then reflex, per a reflex algorithm, to anemia testing.
  • One example may have a celiac panel with the of selecting a celiac panel follow-up.
  • Thyroid panel may algorithm to certain sub tests such as but not limited to T ' 3, T4, or other sub tests related to thyroid function and hormones produced by, or which affect, the thyroid.
  • the back of the lab order form (or other location on the order form) describes what is in each of the panels.
  • This ability to have a lab order form follow a pre-selected algorithm for follow-up testing on micro-sample from the same sample collection session can reduce unnecessary lab visits and unnecessary prescription associated with not having a follow-on order because one needs more tubes of blood are required in order to use traditional dedicated discrete analyzers and each of those analyzers requires its own tube of blood with its own dedicated preparatory reagents and/or dedicated discrete anti-coagulants. Therefore, in a legacy model, the system requires more tubes of blood from the subject and this typically requires a follow-on draw, more visits, and one does not get the diagnosis until later.
  • FIG. 6A A further example of a lab order form (an order sheet) including listings of individual tests, test panels, test groups, and reflex tests according to the methods disclosed herein is provided in Figs. 6A and 6B.
  • tests offered may be grouped together with other tests directed to similar indications or organs (e.g., tests listed under the heading "Thyroid"), or grouped according to general applicability (e.g., tests fisted under the heading "Reproductive Hormones”), listed alphabetically (e.g., tests listed under the headings "Alphabetical Tests (A-H)" and “Alphabetical Tests (H-Z)”), listed according to type of sample (e.g., tests listed under "Stool” and "Urine”), or other groups.
  • A-H Alphahabetical Tests
  • H-Z Alphahabetical Tests
  • a lab order form as disclosed herein may include lines, spaces, boxes, or other locations for entering patient information, clinician information, insurance information, billing information, and may include locations for particulars regarding testing (e.g., whether or not the tests are for a fasting patient; whether or not the test is urgent, or is routine; whether or not the test is to be performed repeatedly, and if so, at what frequency; and so forth).
  • testing e.g., whether or not the tests are for a fasting patient; whether or not the test is urgent, or is routine; whether or not the test is to be performed repeatedly, and if so, at what frequency; and so forth).
  • Clinician information may include clinician name, NPI number, address, and other clinician information.
  • a lab order form as illustrated in Fig. 6A may include a location to fill in a date after which the test order is no longer valid (e.g., "valid until" location).
  • a lab order form may provide a location for a signature (e.g., for the signature of the ordering clinician).
  • a lab order form as disclosed herein may include a Q code, a bar code, or other machine-readable code or feature. Such a machine-readable feature may be effective to more readily enter, store, track, and retrieve patient information.
  • Figure 6A also shows one embodiment of the lab order form wherein algorithm-based testing (designated in phantom by doited rectangles) is shown wherein the option to include is select- ble by the ordering physician.
  • algorithm-based testing designated in phantom by doited rectangles
  • this lab order form with algorithm-based testing differs in that additional testing is offered for tests that traditionally do not mclude an option to order such additional follow-up testing. This may be due in part to limitations of traditional devices preventing such options from being physically possible due to sample size limitations or other reasons.
  • test code and pricing are both listed for each selectable line associated with a test or test panel.
  • Figure 6A also shows that there may be at least four or more algorithm-based tests, wherein at least four categories (common panels, reproductive health, STL other, etc..) each includes at least one algorithm-based test.
  • Figure 6A also shows that there may be at least four or more algorithm-based tests, wherein each includes at least one follow-up test based on an detection technology different from detection technique used in the initial test.
  • Figure 6B shows an emboidment that may optoinaliy include a reverse side on the fab order form.
  • the reverse side shows details regarding assays that are part of in certain combination test panels.
  • Figs. 7A, 7B, and 7C provide yet further examples of an order form having features as disclosed herein.
  • the first page of the order form illustrated in these figures is shown in Fig. 7A, and lists individual tests, test panels, test groupings, and including lists of reflex tests according to methods disclosed herein.
  • A. portion of the second page of the order form illustrated in Fig. 7 is shown in Fig. 7B, and lists panel components and capped price offerings for the list shown in Fig. 7 A.
  • CPT codes are listed for the panels and panel components.
  • a further portion of the order form of Fig. 7, placed on the third page of the order form illustrated in Fig. 7, is shown in Fig. 7C, and lists panel components and capped price offerings for the list shown in Fig. 7A.
  • CPT codes are listed for the panels and panel components.
  • Figs. 8A and 8B provide yet further examples of an order form having features as disclosed herein.
  • the first page of the order form illustrated in these figures is shown in Fig. 8A, and lists individual tests, test panels, test groupings, and including fists of reflex tests according to methods disclosed herein.
  • Fig. 8B shows the second page of the order form illustrated in Fig, 8A, and lists panel components and capped price offerings for (he list shown in Fig. 8A.
  • CPT codes are listed for the panels and panel components.
  • CPT codes are listed for the panels and panel components.
  • the diamond symbols indicate tests for which particular criteria may exist which may limit reimbursement by a subject's insurance.
  • Asterisks indicate tests for which price caps are offered for multiple tests within a panel grouping.
  • the form also indicates that "Ordered reflex tests only performed when medically appropriate. For Medicare and Meidcaid patients physicians should only order those tests that are medically necessary for diagnosis and treatment.”
  • the lab order forms illustrated in Figs. 5A and 513, Figs. 6A and 613, Figs. 7A, 7B, and 7C, and in Figs. 8A and 8B include prices for each test, including reflex tests, and for panels of tests. Providing price information for each test, and for combinations of tests (e.g., test panels) better allows clinicians and patients to understand the costs of tests that are contemplated.
  • Providing reflex test information, including reflex test costs provides benefits by allowing better pl anning of testing, and may result in fewer patient visits being needed for the same number of tests, since a reflex test may be ordered along with an initial test, and so may obviate any need for a subsequent return visit by a subject after the results of an initial test are obtained.
  • reflex test information including reflex test costs, provides benefits in avoiding delay; the delay that may be avoided is the delay which would otherwise occur if a reflex test were ordered only after the results of an initial test were known, since a reflex test may be ordered along with an initial test, and so may eliminate any unnecessary delay.
  • reflex test information including reflex test costs

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Abstract

L'invention concerne des procédés, des dispositifs et des systèmes pour l'analyse d'échantillons biologiques pour déterminer la présence d'un analyte par un dosage initial, et, en fonction des résultats de l'analyse initiale, par un dosage ultérieur. Un échantillon peut être dilué. Une partie d'un échantillon peut être conservée pour être utilisée dans un tel dosage ultérieur. Les résultats du dosage initial peuvent être utilisés pour déterminer, par exemple, un ou plusieurs éléments parmi : si un dosage ultérieur est effectué ou non ; quel dosage ultérieur est effectué ; le procédé de réalisation d'un dosage ultérieur ; l'ordre de réalisation d'une séquence de dosages ultérieurs ; les étapes, ou l'ordre des étapes, effectuées dans un dosage ultérieur ; la synchronisation de la réalisation d'un dosage ultérieur ; le choix d'un réactif utilisé dans un dosage ultérieur ; procédé de détection utilisé dans un dosage ultérieur ; et d'autres détails de dosages pouvant être subordonnée aux résultats d'un essai antérieur.
PCT/US2015/035112 2014-06-11 2015-06-10 Procédés, dispositifs et systèmes pour l'analyse d'échantillons WO2015191716A2 (fr)

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CA2951558A CA2951558A1 (fr) 2014-06-11 2015-06-10 Procedes, dispositifs et systemes pour l'analyse d'echantillons
MX2016016234A MX2016016234A (es) 2014-06-11 2015-06-10 Metodos, dispositivos y sistemas para analisis de muestras.
EP15807539.0A EP3155420A4 (fr) 2014-06-11 2015-06-10 Procédés, dispositifs et systèmes pour l'analyse d'échantillons

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US201462011016P 2014-06-11 2014-06-11
US62/011,016 2014-06-11
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108267599A (zh) * 2018-04-08 2018-07-10 中国人民解放军南京军区南京总医院 一种便携式血液检测分析卡

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EP0397424A3 (fr) * 1989-05-08 1991-08-21 Biotrack, Inc. Système d'analyse multiple
WO2006108263A1 (fr) * 2005-04-15 2006-10-19 Thermo Crs Ltd. Procede et systeme de verification d'echantillons
US20070111225A1 (en) * 2005-08-10 2007-05-17 California Institute Of Technology System and method for monitoring an analyte
JP4922682B2 (ja) * 2006-06-29 2012-04-25 シスメックス株式会社 分析装置
NZ584963A (en) * 2007-10-02 2012-11-30 Theranos Inc Modular Point-of-care devices as addressible assay units with tips of assay units having interior to immobilize reagents by capillary action

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108267599A (zh) * 2018-04-08 2018-07-10 中国人民解放军南京军区南京总医院 一种便携式血液检测分析卡

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WO2015191716A3 (fr) 2016-03-17
EP3155420A4 (fr) 2018-03-07

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