WO2013033727A1 - Système de détection reposant sur la chimioluminescence - Google Patents

Système de détection reposant sur la chimioluminescence Download PDF

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Publication number
WO2013033727A1
WO2013033727A1 PCT/US2012/053705 US2012053705W WO2013033727A1 WO 2013033727 A1 WO2013033727 A1 WO 2013033727A1 US 2012053705 W US2012053705 W US 2012053705W WO 2013033727 A1 WO2013033727 A1 WO 2013033727A1
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WO
WIPO (PCT)
Prior art keywords
cassette
region
electromagnetic radiation
handling device
sample
Prior art date
Application number
PCT/US2012/053705
Other languages
English (en)
Inventor
Charles Franklin Barnett
Ashan R. PERERA
Matthew Huddleston
Jack Bryan Stubbs
Jerome E. Reckelhoff
Jeffrey S. Trapp
Original Assignee
Nanodetection Technology, 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 Nanodetection Technology, Inc. filed Critical Nanodetection Technology, Inc.
Publication of WO2013033727A1 publication Critical patent/WO2013033727A1/fr

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54386—Analytical elements
    • G01N33/54387—Immunochromatographic test strips
    • G01N33/54388—Immunochromatographic test strips based on lateral flow

Definitions

  • This invention generally relates to the detection of biologically relevant materials in a sample.
  • the invention teaches a cassette device for determining the presence or absence of one or more substances in a sample, including: a lateral flow strip, including (1) a first region for accepting a liquid sample, (2) a second region for accepting a luminous reagent, (3) a third region including an enzyme conjugate, (4) a fourth region including a reaction zone, wherein the reaction zone includes a control region and a substance binding region, and wherein (i) the first through fourth regions are in fluid communication, (ii) the control region includes a means for trapping the enzyme conjugate, (iii) the enzyme conjugate is capable of binding with the substance and reacting with a luminous reagent, thereby generating electromagnetic radiation, and (iv) the substance binding region includes a means for binding the substance.
  • the cassette device includes a housing, including an inferior component and a superior component configured to mechanically interact with one another, wherein (1) the superior component includes a first well for adding a sample and a second well for adding a luminous reagent; (2) the inferior component includes a region upon which the lateral flow strip rests; and (3) the first and second wells are in fluid communication with the lateral flow strip. In some embodiments, one or more of the first through fourth regions are partially or completely overlapping.
  • the enzyme conjugate includes an oxidizing agent selected from the group consisting of horseradish peroxidase (HRP), alkaline phosphatase, xanthine oxidase, beta-D galactosidase, glucose oxidase and glucose 6-phosphate dehydrogenase.
  • the luminous reagent is selected from the group consisting of luminol, fluorescein and diphenyl oxalate.
  • the cassette device further includes an aperture on the superior side of the housing for allowing the electromagnetic radiation generated at the substance binding region and/or the control region to escape from the device.
  • the cassette device further includes one or more optical components integrated on the superior side of the cassette, above the control region and/or the substance binding region.
  • one or more of the optical components have a design selected from the group consisting of: classical, spherical, non-spherical geometric optics, fiber optics, fused fiber optics, internal reflection optical channels and combinations thereof.
  • one or more of the optical components are configured to transmit the electromagnetic radiation produced in the control region and/or substance binding region of the cassette.
  • the inferior side of the housing includes an indentation configured to mechanically interact with a complimentary structure on a cassette handling device.
  • the invention teaches a cassette device for determining the presence or absence of one or more substances in a sample, including: a lateral flow strip, including (1) a first region for accepting a liquid sample, (2) a second region for accepting a luminous reagent, (3) a third region comprising a reaction zone, wherein the reaction zone includes a control region and a substance binding region, and wherein (i) the first through third regions are in fluid communication, (ii) the control region includes a means for trapping an enzyme conjugate, (iii) the enzyme conjugate is capable of binding with the substance and reacting with a luminous reagent, thereby generating electromagnetic radiation, and (iv) the substance binding region includes a means for binding the substance.
  • the cassette device includes a housing, including an inferior component and a superior component configured to mechanically interact with one another, wherein (1) the superior component includes a first well for adding a sample and a second well for adding a luminous reagent; (2) the inferior component includes a region upon which the lateral flow strip rests; and (3) the first and second wells are in fluid communication with the lateral flow strip. In some embodiments, one or more of the first through third regions are partially or completely overlapping.
  • the enzyme conjugate includes an oxidizing agent selected from the group consisting of horseradish peroxidase (HRP), alkaline phosphatase, xanthine oxidase, beta-D galactosidase, glucose oxidase and glucose 6-phosphate dehydrogenase.
  • the luminous reagent is selected from the group consisting of luminol, fluorescein and diphenyl oxalate.
  • the cassette device further includes an aperture on the superior side of the housing for allowing the electromagnetic radiation generated at the substance binding region and/or the control region to escape from the device.
  • the cassette device further includes one or more optical components integrated on the superior side of the cassette, above the control region and/or the substance binding region.
  • one or more of the optical components have a design selected from the group consisting of: classical, spherical, non- spherical geometric optics, fiber optics, fused fiber optics, internal reflection optical channels and combinations thereof.
  • one or more of the optical components are configured to transmit the electromagnetic radiation produced in the control region and/or substance binding region of the cassette.
  • the inferior side of the housing includes an indentation configured to mechanically interact with a complimentary structure on a cassette handling device.
  • the invention teaches a cassette device for determining the presence or absence of one or more substances in a sample, including a housing, wherein the housing includes an inferior component and a superior component configured to mechanically interact with one another; wherein (1) the superior component includes a first well for adding a sample and a second well for adding a luminous reagent; (2) the inferior component includes a region wherein a fluidic system is contained; and (3) the first and second wells are in fluid communication with the fluidic system; and the fluidic system includes (1) a first region for accepting the liquid sample (2) a second region for accepting a luminous reagent (3) a third region including a reaction zone wherein the reaction zone includes a control region and a substance binding region, wherein the first through third regions are in fluid communication; and wherein if a target substance is present in the sample, electromagnetic radiation is produced.
  • the fluidic system further includes a fourth region for storing the reagent and a fifth region for mixing the reagent; and the first through fifth regions are in fluid communication.
  • the luminous reagent is selected from the group consisting of luminol, fluorescein and diphenyl oxalate.
  • the cassette device further includes an aperture on the superior side of the housing for allowing the electromagnetic radiation generated at the substance binding region and/or the control region to escape from the device.
  • the cassette device includes one or more optical components integrated on the superior side of the cassette, above the control region and/or the substance binding region.
  • one or more of the optical components have a design selected from the group consisting of: classical, spherical, non-spherical geometric optics, fiber optics, fused fiber optics, internal reflection optical channels and combinations thereof.
  • one or more of the optical components are configured to transmit the electromagnetic radiation produced in the control region and/or substance binding region of the cassette.
  • the underside of the inferior side of the housing includes an indentation configured to mechanically interact with a complimentary structure on a cassette handling device.
  • the cassette device further includes one or more additional indentations configured to mechanically interact with one or more additional complimentary structures on the cassette handling device.
  • one or more of the indentations are hemispherical.
  • the cassette device further includes a means for identifying the cassette device, selected from the group consisting of: RF, optical, barcode, QR barcode, one or more electrical contacts capable of conducting an electrical signal and combinations thereof.
  • the invention teaches a cassette handling device, including a cassette acceptor, including a superior side and an inferior side; a loading mechanism for guiding a cassette into a predetermined position; a cam gear located on the inferior side of the cassette acceptor and configured to provide a continuous feed of the cassette into the predetermined position upon insertion into the cassette handling device; and an aperture configured on the superior side of the cassette acceptor.
  • the cassette handling device further includes a base, a spring, and a projection
  • the spring includes a first end and a second end and the projection includes a first end and a second end; the first end of the spring mechanically interacts with a base of the cassette handling device, and the second end of the spring is attached to the first end of the projection; the second end of the projection is configured to mechanically interact with a complimentary region of the cassette when the cassette is in the predetermined position, thereby stabilizing the cassette in the predetermined position.
  • the projection is spherical.
  • the cassette handling device further includes an alignment grid, wherein the alignment grid includes a first side, a second side and a plurality of holes, and wherein a portion of the alignment grid is aligned with the aperture of the cassette handling device.
  • the invention teaches an electromagnetic radiation detection device, including: (a) a housing, including an aperture; (b) a cassette handling device, including a cassette acceptor, including a superior side and an inferior side; a loading mechanism for guiding a cassette into a predetermined position; a cam gear located on the inferior side of the cassette acceptor and configured to provide a continuous feed of the cassette into the predetermined position upon insertion into the cassette handling device; and an aperture configured on the superior side of the cassette acceptor; wherein the cassette handling device is configured to fit into the housing and wherein at least a portion of the aperture of the housing is configured to align with at least a portion of the aperture of the cassette handling device; and (c) an electromagnetic radiation detector.
  • the cassette handling device further includes a spring, a projection and a base, wherein the spring includes a first end and a second end and the projection includes a first end and a second end; the first end of the spring mechanically interacts with the base of the cassette handling device, and the second end of the spring is attached to the first end of the projection; the second end of the projection is configured to mechanically interact with a complimentary region of the cassette when the cassette is in the predetermined position, thereby stabilizing the cassette in the predetermined position.
  • the projection is spherical.
  • the cassette handling device further includes an alignment grid
  • the alignment grid includes a first side, a second side and a plurality of holes
  • one or more of the holes of the alignment grid are aligned with the aperture of the cassette handling device on the first side and the detector on the second side.
  • the electromagnetic radiation detection device includes one or more components selected from the group consisting of: a CMOS chip with a discrete detection region, a charge-coupled device, an electro-optical sensor, a photodetector, a photodiode, a photomultiplier tube, a single-photon avalanche diode and a visible light photon counter.
  • the electromagnetic radiation detection device further includes a display, wherein the display is capable of displaying data regarding the sample, based upon the electromagnetic radiation detected.
  • the invention teaches a system for detecting the presence or absence of a substance in a sample; the system includes: (a) a cassette, including: a lateral flow strip, including (1) a first region for accepting a liquid sample, (2) a second region for accepting a luminous reagent, (3) a third region including an enzyme conjugate, (4) a fourth region including a reaction zone, wherein the reaction zone includes a control region and a substance binding region, and wherein (i) the first through fourth regions are in fluid communication, (ii) the control region includes a means for trapping the enzyme conjugate, (iii) the enzyme conjugate is capable of binding with the substance and reacting with a luminous reagent, and (iv) the substance binding region includes a means for binding the substance; and a housing, including an inferior component and a superior component configured to mechanically interact with one another, wherein (1) the superior component includes a first well for adding a sample and a second well for adding a luminous reagent; (2) the inferior component
  • the cassette handling device further includes a spring, a projection and a base, wherein the spring includes a first end and a second end and the projection includes a first end and a second end; the first end of the spring mechanically interacts with a base of the cassette handling device, and the second end of the spring is attached to the first end of the projection; the second end of the projection is configured to mechanically interact with a complimentary region of the cassette, when the cassette is in the predetermined position, thereby stabilizing the cassette in the predetermined position.
  • the projection is spherical.
  • the cassette handling device further includes an alignment grid; the alignment grid includes a first side, a second side and a plurality of holes, and one or more of the holes of the alignment grid are aligned with the aperture of the cassette handling device on the first side and the detector on the second side.
  • the electromagnetic radiation detection device of the system includes one or more components selected from the group consisting of: a CMOS chip with a discrete detection region, a charge-coupled device, an electro-optical sensor, a photodetector, a photodiode, a photomultiplier tube, a single-photon avalanche diode and a visible light photon counter.
  • the system further includes a display, wherein the display is capable of displaying data regarding the sample, based upon the electromagnetic radiation detected.
  • the enzyme conjugate includes an oxidizing agent selected from the group consisting of horseradish peroxidase (HRP), alkaline phosphatase, xanthine oxidase, beta-D galactosidase, glucose oxidase and glucose 6-phosphate dehydrogenase.
  • HRP horseradish peroxidase
  • alkaline phosphatase alkaline phosphatase
  • xanthine oxidase beta-D galactosidase
  • glucose oxidase glucose 6-phosphate dehydrogenase
  • one or more of the first through fourth regions of the cassette are partially or completely overlapping.
  • the luminous reagent is selected from the group consisting of luminol, fluorescein and diphenyloxalate.
  • the cassette further includes an aperture on the superior side of the cassette housing for allowing the electromagnetic radiation generated at the substance binding region and/or the control region to escape from the device.
  • the cassette of the system further includes one or more optical components integrated on the superior side of the cassette, above the control region and/or the substance binding region.
  • one or more of the optical components have a design selected from the group consisting of: classical, spherical, non-spherical geometric optics, fiber optics, fused fiber optics, internal reflection optical channels and combinations thereof.
  • one or more of the optical components are configured to transmit the electromagnetic radiation produced in the control region and/or substance binding region of the cassette.
  • the inferior side of the housing of the cassette includes an indentation configured to mechanically interact with a complimentary structure on the cassette handling device.
  • the cassette includes one or more additional indentations configured to mechanically interact with one or more additional complimentary structures on the cassette handling device.
  • one or more of the indentations are hemispherical.
  • the cassette of the system further includes a means for identifying the cassette, selected from the group consisting of: RF, optical, barcode, QR barcode, one or more electrical contacts capable of conducting an electrical signal and combinations thereof.
  • the invention teaches a method for analyzing a sample, including: providing a sample; and utilizing (1) any one of the aforementioned cassette devices, (2) any one of the aforementioned cassette handling devices, (3) any one of the aforementioned electromagnetic radiation detection devices; or (4) any one of the aforementioned systems to analyze the sample.
  • the method further includes detecting the presence or absence of a substance in the sample.
  • Figure 1A depicts, in accordance with an embodiment of the invention, a cassette 100 with an aperture 101, a housing 102 and a well 103.
  • Figure IB depicts, in accordance with an embodiment of the invention a cassette 200 with integrated optics 201, a cassette housing 202 and a well 203.
  • Figure 2A depicts, in accordance with an embodiment of the invention, the top half of a cassette device 400, with integrated optics 401, a housing 402 and the top side of a region 403 configured to contain a means for cassette identification.
  • Figure 2B depicts, in accordance with an embodiment of the invention, the bottom half of a cassette device 400 with a housing 402, a lateral flow strip 405, a region 406 of the lateral flow strip for receiving a fluid from the well 404 (depicted in 2A) and a region 407 configured to contain a means for cassette identification.
  • Figure 2C depicts, in accordance with an embodiment of the invention, the underside of the lower half of the cassette device 400 with a housing 402 and spherical indentations 408.
  • Figure 2D depicts, in accordance with an embodiment of the invention a cassette device 500 with a region 501 to accommodate a means for identification, rows of blocked lenses 502, lenses 503 and a well 504.
  • Figure 3 depicts, in accordance with an embodiment of the invention, a ray trace diagram.
  • Figure 4A depicts, in accordance with an embodiment of the invention, a cassette handler 600 with a cassette receiving component 601, a mechanism 602 for introducing a cassette into the cassette handler 600, a cam gear 603 and an aperture 604.
  • Figure 4B depicts, in accordance with an embodiment of the invention, a cut out view of a cassette handler 600 in which the base 605, spring 606 and projection 607 components can be seen.
  • Figure 5 A depicts, in accordance with an embodiment of the invention, a head on view of a system for detecting electromagnetic radiation on a cassette 701.
  • the system includes a cassette handler 700, a cassette 701 with integrated lenses 702, an alignment grid 703 with holes 704, an electromagnetic radiation detector 705 and a thermoelectric cooler 706, which can be optionally included in the system.
  • Figure 5A also shows the integration of signal processing components (depicted as box “A"), and display and user interface components (depicted as box “B”).
  • Figure 5B depicts a view in which each of the components are separated.
  • Figure 6A depicts, in accordance with an embodiment of the invention a housing 800 configured to accommodate an array of lenses in the individual sockets 801.
  • Figure 6B depicts a photodiode array 1001 of an electromagnetic radiation detection device 1000 interacting with probes 901 of a device 900.
  • Figure 7 depicts, in accordance with an embodiment of the invention, an array 1101 of a detection device 1100.
  • Figure 8 A depicts, in accordance with an embodiment of the invention, an electromagnetic radiation detection system, including a cassette device 400 inserted into a cassette handler 600, with an electromagnetic radiation detector 1201 resting above the region of the cassette 400 in which electromagnetic radiation is generated as a result of a chemical reaction.
  • the device 1200 has been fitted with a display screen 1201 and the cartridge handler is covered, but the cassette 400 inserted therein can still be seen.
  • “Mammal” as used herein refers to a member of the class Mammalia, including, without limitation, humans as well as nonhuman primates such as chimpanzees and other apes and monkey species; farm animals such as cattle, sheep, pigs, goats and horses; domestic mammals such as dogs and cats; laboratory animals including rodents such as mice, rats and guinea pigs, and the like.
  • the term does not denote a particular age or sex. Thus, newborn subjects and infant subjects, as well as fetuses, whether male or female, are intended to be included within the scope of this term.
  • the terms “detect,” “detecting,” or “detection” may describe either the general act of discovering or discerning or the specific observation of a detectably labeled composition, or a composition that doesn't require labeling for detection.
  • the numbers expressing quantities of ingredients, properties such reaction conditions, and so forth, used to describe and claim certain embodiments of the application are to be understood as being modified in some instances by the term "about.” Accordingly, in some embodiments, the numerical parameters set forth in the written description and attached claims are approximations that can vary depending upon the desired properties sought to be obtained by a particular embodiment. In some embodiments, the numerical parameters should be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. Notwithstanding that the numerical ranges and parameters setting forth the broad scope of some embodiments of the application are approximations, the numerical values set forth in the specific examples are reported as precisely as practicable.
  • Polypeptide and “protein” are used interchangeably herein and include all polypeptides as described below.
  • the basic structure of polypeptides is well known and has been described in innumerable textbooks and other publications in the art.
  • the term is used herein to refer to any peptide or protein comprising two or more amino acids joined to each other in a linear chain by peptide bonds.
  • the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types.
  • polypeptides often contain amino acids other than the 20 amino acids commonly referred to as the 20 naturally occurring amino acids, and that many amino acids, including the terminal amino acids, may be modified in a given polypeptide, either by natural processes, such as processing and other post-translational modifications, but also by chemical modification techniques which are well known in the art. Even the common modifications that occur naturally in polypeptides are too numerous to list exhaustively here, but they are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature, and they are well known to those of skill in the art.
  • polypeptides used in conjunction with the present invention are, to name an illustrative few, acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphatidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-R A mediated addition of amino acids to proteins such as arginylation, and ubiquitination.
  • the substance detected by one or more of the inventive devices and methods can include, but is in no way limited to a protein, a peptide, a virus (or portion thereof), a genetically engineered virus (or portion thereof), a bacterium (or portion thereof), a genetically engineered bacterium (or portion thereof), a fungi (or portion thereof), a genetically engineered fungi (or portion thereof), a drug, a drug metabolite, a lipid, a fatty acid, a nucleic acid, a polysaccharide, a chemical, an enzyme, a hormone, a toxin, a chemical weapon, a biological weapon, or combinations thereof.
  • the substance is indirectly detected by detecting an antibody to the substance, generated by an individual.
  • the individual is an animal.
  • the individual is a mammal.
  • the individual is a human.
  • Non-limiting examples of sources of samples that can be analyzed using the present invention include an animal, a human, a plant, an insect, a body of water, sewage or a culture.
  • the sample that can be used in conjunction with one or more of the inventive devices and methods can include, but are in no way limited to, blood, serum, nasal fluid, urine, sweat, plasma, semen, cerebrospinal fluid, tears, pus, amniotic fluid, saliva, lung aspirate, gastrointestinal contents, vaginal discharge, urethral discharge, chorionic villi specimens, skin epithelials, genitalia epithelials, gum epithelials, throat epithelials, hair or sputum.
  • the sample is obtained from an animal.
  • the sample is obtained from a mammal.
  • the sample is obtained from a human.
  • antibody encompasses all antibody molecules, obtained from both polyclonal and monoclonal preparations.
  • the term also includes hybrid (chimeric) antibody molecules, and any functional fragments obtained from such molecules, wherein such fragments retain immunological binding properties of the parent antibody molecule.
  • the present invention relates to systems, methods and devices for detecting one or more biologically relevant substances in a sample.
  • the invention teaches a cassette device for determining the presence or absence of one or more substances in a sample.
  • the cassette device includes a lateral flow strip.
  • the lateral flow strip includes a region for accepting a liquid sample and a region for accepting a luminous reagent.
  • the lateral flow strip also includes a region containing an enzyme conjugate.
  • the lateral flow strip also includes a region that serves as a reaction zone.
  • the reaction zone includes a control region and a substance binding region, and all of the regions of the lateral flow strip are in fluid communication.
  • the control region includes a means for trapping the enzyme conjugate.
  • the means for trapping the enzyme conjugate is an antibody.
  • the enzyme conjugate is capable of binding with a target substance of the sample and reacting with a luminous reagent, thereby generating electromagnetic radiation.
  • the enzyme conjugate includes an antibody to facilitate binding with the target substance in a sample.
  • the substance binding region of the lateral flow strip includes a means for binding the target substance of the sample.
  • the means for binding the target substance is an antibody.
  • the cassette device 400 includes a housing 402.
  • the cassette device includes an inferior component ( Figure 2B) and a superior component (Figure 2A) configured to mechanically interact with one another.
  • the superior component ( Figure 2A) includes a well 404.
  • the cassette device includes one or more additional wells. In each of the aforementioned embodiments, the wells are in fluid communication with a region of a lateral flow strip.
  • the inferior component ( Figure 2B) comprises a region upon which the lateral flow strip 405 rests.
  • the lateral flow strips of the devices described herein include a negative control region.
  • the negative control region includes an antibody that is not specific for the enzyme conjugate.
  • the negative control region can be used to determine the level of non-specific binding of the enzyme conjugate. Once the level of non-specific binding of the enzyme conjugate is determined, that contribution to a positive signal can be determined and accounted for, in order to provide a more accurate reading of the assay results.
  • the secondary antibody labeled with the enzyme may be added separately and included in a sample preparation bottle.
  • the enzyme conjugate used in conjunction with the inventive cassette devices described above includes an oxidizing agent selected from the group consisting of horseradish peroxidase (HRP), alkaline phosphatase, xanthine oxidase, beta-D galactosidase, glucose oxidase and glucose 6-phosphate dehydrogenase.
  • HRP horseradish peroxidase
  • alkaline phosphatase alkaline phosphatase
  • xanthine oxidase beta-D galactosidase
  • glucose oxidase glucose 6-phosphate dehydrogenase
  • the luminous reagent used in conjunction with the inventive cassette devices described above is selected from the group consisting of luminol, fluorescein and diphenyl oxalate.
  • luminol a group consisting of luminol, fluorescein and diphenyl oxalate.
  • alternative luminous reagents comparable to those specifically described herein, could also be used with the inventive devices described herein.
  • the enzyme conjugate used with the inventive cassette device is horseradish peroxidase (HRP) and the luminous reagent is luminol.
  • HRP horseradish peroxidase
  • the lateral flow strip can be made of materials including glass fiber and/or nitrocellulose.
  • the lateral flow strip can also include nylon-based membranes.
  • a conjugate pad, onto which the enzyme conjugate is dried is also used.
  • the conjugate pad is integrated within a sample pad, and when re- hydration of the conjugate occurs by introducing a liquid sample onto the sample pad, interaction between the enzyme conjugate and the targeted substance of the sample begins.
  • the cassette device 100 includes an aperture 101 on the superior side of the housing 102 for allowing the electromagnetic radiation generated at the substance binding region and/or the control region to escape from the cassette device 100.
  • the cassette device 200 further includes one or more optical components 201 integrated on the superior side of the cassette 200, above the control region and/or the substance binding region.
  • the optical components integrated into the cassette device have a design selected from the group consisting of: classical, spherical, non-spherical geometric optics, fiber optics, fused fiber optics, internal reflection optical channels and combinations thereof.
  • the optics can be configured as an optical array 201, designed to transmit the electromagnetic radiation produced in the control region and/or substance binding region of the cassette 200.
  • one or more of the optical components (including a row of optical components) of the cassettes described herein can be blocked.
  • Figure 2D depicts an embodiment in which a row of lenses 503 are blocked 502.
  • the underside of the inferior side of the cassette housing 402 of the cassette 400 includes one or more indentations 408 configured to mechanically interact with complimentary structures on a cassette handling device.
  • These indentations are especially well suited to interact with complimentary structures on the appropriately configured cassette handling devices described herein, such as those depicted in Figures 4A and 4B, containing complimentary structures 607.
  • one or more of the indentations 408 on the cassette 400 are hemispherical, as depicted in Figure 1C.
  • One of skill in the art would readily appreciate that one or more of the indentations could be of a different shape, so long as they are configured to mechanically interact with one or more complimentary regions on a cassette handling device.
  • the indentations described above can be configured on any cassette described herein.
  • the cassette includes a means by which the cassette device can be identified.
  • the means by which the cassette device can be identified can include, but are in no way limited to: RF, optical, barcode, QR barcode, one or more electrical contacts capable of conducting an electrical signal and combinations thereof.
  • Figures 2A and 2B depict a region (403 and 407, respectively) of a cassette device 400, in which a means by which the cassette can be identified can be situated.
  • a means by which the cassette can be identified can be situated.
  • the invention teaches a cassette device for determining the presence or absence of one or more substances in a sample without the use of a lateral flow strip.
  • the cassette device includes a housing, wherein the housing includes an inferior component and a superior component configured to mechanically interact with one another, wherein (1) the superior component includes a well for adding a sample and a well for adding a luminous reagent; (2) the inferior component includes a region wherein a fluidic system is contained; and (3) the wells are in fluid communication with the fluidic system.
  • the fluidic system includes (1) a first region for accepting the liquid sample (2) a second region for accepting a luminous reagent and (3) one or more additional regions including a reaction zone.
  • the reaction zone includes a control region and a substance binding region, and all of the aforementioned regions are in fluid communication.
  • the fluidic system further includes an additional region for reagent storage, as well as a region for reagent mixing, and all of the aforementioned regions are in fluid communication.
  • the device is configured in such a manner that if one or more specific target substances are present in a sample applied to the device, electromagnetic radiation will be produced as a result of a chemical reaction.
  • the enzyme conjugate used in conjunction with the above- described cassette devices, without a lateral flow strip includes an oxidizing agent selected from the group consisting of horseradish peroxidase (HRP), alkaline phosphatase, xanthine oxidase, beta-D galactosidase, glucose oxidase and glucose 6-phosphate dehydrogenase.
  • HRP horseradish peroxidase
  • alkaline phosphatase alkaline phosphatase
  • xanthine oxidase beta-D galactosidase
  • glucose oxidase glucose 6-phosphate dehydrogenase
  • the luminous reagent used in conjunction with the above- described inventive devices is selected from the group consisting of luminol, fluorescein and diphenyl oxalate.
  • luminol a group consisting of luminol, fluorescein and diphenyl oxalate.
  • alternative luminous reagents comparable to those specifically described herein with respect to their interaction with oxidizing agents, could be used with the inventive devices described herein.
  • the enzyme conjugate used in conjunction with the inventive cassette devices described above is HRP
  • the luminous reagent is luminol
  • the cassette device further includes an aperture on the superior side of the housing for allowing the electromagnetic radiation generated at the substance binding region and/or the control region to escape from the device.
  • the cassette device further includes one or more optical components integrated on the superior side of the cassette, above the control region and/or the substance binding region.
  • the optical components integrated into the cassette device have a design selected from the group consisting of: classical, spherical, non-spherical geometric optics, fiber optics, fused fiber optics, internal reflection optical channels and combinations thereof.
  • One or more of the optical components can be configured to transmit the electromagnetic radiation produced in the control region and/or substance binding region of the cassette.
  • the underside of the inferior side of the cassette housing includes an indentation, configured to mechanically interact with a complimentary structure on a cassette handling device.
  • the cassette includes one or more additional indentations configured to mechanically interact with one or more additional complimentary structures on the cassette handling device.
  • one or more of the indentations on the cassette are hemispherical.
  • One of skill in the art would readily appreciate that one or more of the indentations could be of a non-hemispherical shape, so long as it is configured to be complimentary to a cassette handling device described herein.
  • the cassette also includes a means by which the cassette device can be identified.
  • the means by which the cassette device can be identified can include, but are in no way limited to: RF, optical, barcode, QR barcode, one or more electrical contacts capable of conducting an electrical signal and combinations thereof.
  • the electromagnetic radiation generated by an assay conducted using any of the various cassette devices described herein could be detected using any of a number of appropriately configured detection devices, including those described herein.
  • the invention teaches a cassette handling device.
  • the cassette handling device 600 includes a cassette acceptor 601, including a superior side and an inferior side; a loading mechanism 602 for guiding a cassette 400 ( Figures 2A and 2B) into a predetermined position within the cassette handling device 600; a cam gear 603 located on the inferior side of the cassette acceptor 601 and configured to provide a continuous feed of the cassette 400 into the predetermined position upon insertion into the cassette handling device 600; and an aperture 604 configured on the superior side of the cassette acceptor 601.
  • the cassette handling device 600 also includes a base 605, a spring 606, and a projection 607.
  • the spring 606 includes a first end and a second end and the projection includes a first end and a second end.
  • the first end of the spring 606 mechanically interacts with a base 605 of the cassette handling device 600, and the second end of the spring 606 is attached to the first end of the projection 607.
  • the second end of the projection is configured to mechanically interact with a complimentary region of a cassette 400, when the cassette is in the predetermined position, thereby stabilizing the cassette 400 in the predetermined position.
  • the projection 607 is spherical, as depicted in Figure 4B.
  • the cassette handling device includes a hinged door, configured to substantially block the opening of the cassette acceptor and to articulate into the cassette acceptor when a cassette is inserted into the cassette handling device.
  • the hinge is configured across the top of the cassette handler.
  • the hinged door is configured to close behind a cassette inserted into the cassette handling device.
  • the cassette handling device also includes an alignment grid 703, including a first side, a second side and a plurality of holes 704. In some embodiments, at least a portion of the alignment grid 703 is aligned with at least a portion of the optical components 702 of the cassette 701, as depicted in the head on view of Figures 5 A and 5B.
  • the various cassette handling devices of the present invention can be incorporated within any number of configurations of electromagnetic radiation detection devices, in order to facilitate alignment of the aperture/optics of a cassette with one or more detectors of the electromagnetic radiation detection devices, while simultaneously reducing the entrance of stray light into the area of detection.
  • the invention teaches an electromagnetic radiation detection device, including: a cassette handling device 600, including a cassette acceptor 601, including a superior side and an inferior side; a loading mechanism 602 for guiding a cassette into a predetermined position; a cam gear 603 located on the inferior side of the cassette acceptor 601 and configured to provide a continuous feed of the cassette into the predetermined position upon insertion into the cassette handling device 600; and an aperture 604 configured on the superior side of the cassette acceptor 601; wherein the cassette handling device 600 is configured to fit into the electromagnetic radiation detection device, and wherein at least a portion of the aperture of the cassette handling device 604 and an electromagnetic radiation detector of the electromagnetic radiation detection device are aligned.
  • FIG. 8 A An embodiment of the electromagnetic radiation detection device 1200 is depicted in Figure 8 A.
  • Figure 8 A shows a cassette 400 inserted into a cassette handler 600.
  • Figure 8 A also shows an electromagnetic radiation detector, configured above the region of the cassette in which electromagnetic radiation is produced.
  • the cassette handling device further includes a spring 606, a projection 607 and a base 605, as previously described herein.
  • the cassette handling device further includes an alignment grid 703.
  • the alignment grid 703 includes a first side, a second side, and a plurality of holes 704.
  • one or more of the holes 704 of the alignment grid 703 are aligned with the optics of a cassette device 701 on the first side and one or more regions of the detector 705 on the second side.
  • the aperture of the cassette handling device is configured to accept rows of lenses or an array of lenses from a cassette described herein.
  • the cassette handling device is secured to the electromagnetic radiation detection device by a housing that surrounds the cassette handling device.
  • the housing contains an aperture to that at least partially aligns with the aperture of the cassette handling device on one side and one or more detectors of the electromagnetic radiation detection device on the other.
  • the electromagnetic radiation detection device also includes one or more detection components selected from the group consisting of: a CMOS chip with a discrete detection region, a charge-coupled device, an electro-optical sensor, a photodetector, a photodiode, a photomultiplier tube, a single-photon avalanche diode and a visible light photon counter.
  • the electromagnetic radiation detection device further includes a display 1201, wherein the display is capable of displaying data regarding the sample, based upon the electromagnetic radiation detected.
  • the electromagnetic radiation detection device further includes signal processing components and user interface electronics, depicted as boxes "A" and "B,” respectively, in Figure 5A.
  • the electromagnetic radiation detection device is configured to (1) detect electromagnetic radiation generated as a result of a chemical reaction occurring on a cassette device (2) convert the electromagnetic radiation into an electrical signal, (3) process the electrical signal to determine the identity of one or more substances present or absent in the sample, and (4) communicate whether a substance is present or absent in a sample after the signal is processed.
  • the electromagnetic radiation detection device is further configured to wirelessly communicate the data collected therein to one or more additional devices. The components and various configurations thereof required to carry out these operations are well known to one of skill in the art.
  • the CMOS detector in the present invention functions by allowing independent bias to be applied to each detection area and independent readout of the detected analog signals in the integrated circuit array.
  • the invention teaches a system for detecting the presence or absence of a substance in a sample.
  • the system includes a cassette, including: a lateral flow strip.
  • the lateral flow strip includes a region for accepting a liquid sample, and a region for accepting a luminous reagent.
  • the lateral flow strip also includes a region including an enzyme conjugate.
  • the lateral flow strip includes a region including a reaction zone, wherein the reaction zone includes a control region and a substance binding region.
  • each of the aforementioned regions are in fluid communication
  • the control region includes a means for trapping the enzyme conjugate
  • the enzyme conjugate is capable of binding with the substance and reacting with a luminous reagent
  • the substance binding region includes a means for binding the target substance of the sample.
  • the control region and substance binding region function through the use of antibodies as described herein in the description of the cassette device.
  • the remaining regions are in fluid communication with one another.
  • the cassette includes a housing, including an inferior component and a superior component, configured to mechanically interact with one another, wherein (1) the superior component includes a first well for adding a sample and a second well for adding a luminous reagent; (2) the inferior component includes a region upon which the lateral flow strip rests; and (3) the first and second wells are in fluid communication with the lateral flow strip.
  • the cassette also includes indentations for communicating with a cassette handler, as previously described. Any of the alternative configurations of the cassette devices (with or without integrated optics), described in the description of the cassette devices above, can be used in the inventive system.
  • the system further includes a cassette handling device 600, including: a cassette acceptor 601, including a superior side and an inferior side; a loading mechanism 602 for guiding a cassette 400 into a predetermined position; a cam gear 603 located on the inferior side of the cassette acceptor 601 and configured to provide a continuous feed of the cassette 400 into the predetermined position, upon insertion into the cassette handling device 600; and an aperture 604 configured on the superior side of the cassette acceptor 601.
  • the cassette handling device 600 further includes one or more spring 606 and projection 607 components, as previously described herein.
  • the cassette handling device includes an alignment grid, as previously described herein.
  • the system further includes an electromagnetic radiation detector.
  • the electromagnetic radiation detection device of the system includes one or more components selected from the group consisting of: a CMOS chip with a discrete detection region, a charge-coupled device, an electro-optical sensor, a photodetector, a photodiode, a photomultiplier tube, a single-photon avalanche diode and a visible light photon counter.
  • the system also includes a display 1201, wherein the display is capable of displaying data regarding the sample, based upon the electromagnetic radiation generated on a cassette device described herein.
  • the system also includes signal processing components, user interface components and communication components, as described herein.
  • the system is configured to communicate wirelessly with one or more additional devices.
  • the systems and devices of the invention include a power source.
  • the electromagnetic radiation detection device is configured to (1) detect electromagnetic radiation generated as a result of a chemical reaction occurring on a cassette device (2) convert the electromagnetic radiation into an electrical signal, (3) process the electrical signal and thereby determine the presence or absence of one or more target substances in a sample applied to the cassette device, and (4) communicate whether one or more substances are present or absent in a sample after the signal is processed, by generating data visible on a display of the device.
  • the invention teaches a method for analyzing a sample, including: providing a sample, and applying the sample to a cassette device described herein, through one of the sample wells described herein.
  • the invention further includes adding a luminous reagent to a cassette device, through one of the wells of the cassette device described herein and suited therefore.
  • the luminous reagent is added to the sample and then the luminous reagent and sample mixture is added to a single well in a cassette device containing an enzyme conjugate.
  • the luminous reagent and sample are added separately through a single well in a cassette device described herein.
  • the luminous reagent and sample are added to separate wells in a cassette device described herein.
  • the invention teaches a method, including inserting a cassette device of the present invention into a cassette handling of the present invention.
  • the invention teaches a method for analyzing a sample, including detecting electromagnetic radiation emitted from a cassette device of the invention using an electromagnetic radiation detection device of the invention.
  • the invention teaches detecting the presence or absence of one or more substances in a sample, utilizing any of the cassette, cassette handler or electromagnetic radiation detection devices of the claimed invention, according to the methods described herein.
  • Table 1 provides a non-comprehensive supplemental list of pathogens that may be detected using the inventive devices, systems and methods disclosed herein.
  • a swab of an infected skin lesion containing MRSA is obtained from a patient.
  • the portion of the swab containing the sample is broken off and introduced into a container with 1 ml of saline, and vortexed for 1 minute to distribute the sample into the saline.
  • 100 ⁇ of the saline containing the swabbed sample is introduced into a sample well of a MRSA test cassette device.
  • the liquid sample will flow through the sample well and onto a lateral flow strip embedded within the device.
  • the liquid sample will then re-hydrate a portion of the lateral flow strip containing a conjugate made of horseradish peroxidase (HRP) and an antibody specific for MRSA.
  • HRP horseradish peroxidase
  • conjugate will bind to MRSA present in the sample, and both the bound and unbound conjugate will then migrate along the lateral flow strip to a reaction zone containing a control region and a testing region.
  • the control region contains a stripe of antibody specific for HRP and the testing region contains a stripe of an antibody specific for MRSA.
  • Unbound conjugate will bind at the control region, whereas conjugate bound to MRSA will bind at the test region.
  • Luminol is then introduced into a luminol well of the test cassette device, and flows through the luminol well and onto the lateral flow strip. The luminol migrates along the device to the control region and testing region, where it reacts with the HRP trapped in both regions and produces electromagnetic radiation.
  • the electromagnetic radiation is transmitted through rows of lenses integrated onto the top portion of the device and positioned directly above the control and testing regions.
  • the cassette is then inserted into the cassette acceptor of a cassette handling device, which is located within an electromagnetic radiation detection device.
  • a cam gear of the cassette handling device guides the cassette into a position in the cassette handling device that aligns the lenses of the cassette with electromagnetic radiation detectors of the detection device and reduces the entry of stray light into the detectors, which are positioned directly above the cassette handling device.
  • the cassette is then stabilized in that position by the mechanical interaction of four hemispherical indentations (located approximately at the corners of the underside of the cassette) with four spring mounted spherical projections of the cassette handler.
  • the electromagnetic radiation produced in the control and testing regions of the cassette is then detected by the detectors, and an electrical signal is produced in response to the detection.
  • the electrical signals are processed by a processor located within the electromagnetic radiation detection device, and then a signal indicating the presence of MRS A is generated and communicated to a display of the electromagnetic detection device, where it can be viewed by a user.

Abstract

La présente invention concerne des dispositifs, des systèmes et des procédés permettant de détecter par chimioluminescence une ou plusieurs substances dans un échantillon. L'intégration d'un élément optique dans un dispositif à cassette et/ou la manière dont un dispositif à cassette interagit avec un dispositif de détection de rayonnement électromagnétique peuvent/peut présenter des avantages significatifs par rapport aux dispositifs classiques.
PCT/US2012/053705 2011-09-02 2012-09-04 Système de détection reposant sur la chimioluminescence WO2013033727A1 (fr)

Applications Claiming Priority (12)

Application Number Priority Date Filing Date Title
US201161530817P 2011-09-02 2011-09-02
US201161530849P 2011-09-02 2011-09-02
US201161530846P 2011-09-02 2011-09-02
US201161530823P 2011-09-02 2011-09-02
US201161530828P 2011-09-02 2011-09-02
US201161530759P 2011-09-02 2011-09-02
US61/530,849 2011-09-02
US61/530,846 2011-09-02
US61/530,828 2011-09-02
US61/530,759 2011-09-02
US61/530,817 2011-09-02
US61/530,823 2011-09-02

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WO2017172577A1 (fr) * 2016-03-29 2017-10-05 Nanodetection Technology, Inc. Système de détection par chimiluminescence de staphylococcus aureus résistant à la méthicilline

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US20100267049A1 (en) * 2009-04-15 2010-10-21 Rutter William J Diagnostic devices and related methods

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017172577A1 (fr) * 2016-03-29 2017-10-05 Nanodetection Technology, Inc. Système de détection par chimiluminescence de staphylococcus aureus résistant à la méthicilline
CN108884151A (zh) * 2016-03-29 2018-11-23 纳米检测技术有限公司 用于耐甲氧西林金黄色酿脓葡萄球菌的基于化学发光的检测的系统
US10451623B2 (en) * 2016-03-29 2019-10-22 Nanodetection Technology, Inc. System for chemiluminescence-based detection of methicillin-resistant Staphylococcus aureus

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