WO2013192377A1 - Système de diagnostic portable et ses utilisations - Google Patents

Système de diagnostic portable et ses utilisations Download PDF

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Publication number
WO2013192377A1
WO2013192377A1 PCT/US2013/046714 US2013046714W WO2013192377A1 WO 2013192377 A1 WO2013192377 A1 WO 2013192377A1 US 2013046714 W US2013046714 W US 2013046714W WO 2013192377 A1 WO2013192377 A1 WO 2013192377A1
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WIPO (PCT)
Prior art keywords
sample
light
substrate
reagents
camera
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PCT/US2013/046714
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English (en)
Inventor
Chris N. THORNTON
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Global Biodiagnostics
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Application filed by Global Biodiagnostics filed Critical Global Biodiagnostics
Publication of WO2013192377A1 publication Critical patent/WO2013192377A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N21/03—Cuvette constructions
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N21/03—Cuvette constructions
    • G01N2021/0325—Cells for testing reactions, e.g. containing reagents
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • G01N2021/6439—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" with indicators, stains, dyes, tags, labels, marks

Definitions

  • the present invention relates generally to fields of fluorescent or colorimetric imaging and to the detection of bacteria and diagnosis of associated pathophysical conditions. More specifically, the present invention discloses a portable diagnostic system utilizing a fluorophore or fluorescent reporter for fluorescent or colorimetric detection of bacteria and/or enzymes.
  • the present invention is directed to a system for detecting the presence of bacteria of interest in a sample in real time.
  • the system comprises means for containing the sample in which the bacteria may be present, means for providing an excitation wavelength of light to the sample, means for capturing light emitted from the sample after excitation, and means for analyzing the captured light.
  • the present invention is directed to a related system further comprising means for housing the system.
  • the present invention is further directed to another system for detecting the presence of a pathogenic bacteria in a sample in real time.
  • the system comprises a bio- safe container for a sample, configured to eliminate exposure to infectious bacteria, in which the pathogenic bacteria might be present and a fluorescently-labeled substrate and reagents for an enzyme comprising the pathogenic bacteria and a mixing mechanism configured to isolate the sample from the substrate/reagents during sample collection prior to mixing the same after collection.
  • the system comprises one or more light sources each having an excitation wavelength of light deliverable to the sample.
  • the system comprises a camera with camera lens comprising a mobile device or a mobile computing device and an emission filter disposed in alignment between the camera and sample.
  • the system comprises software configured to provide instructions executable by a processor to enable analysis of an image of fluorescent light captured by the camera via one or more of a comparison of light intensity with a threshold, shape or distribution of light intensity or an increase in intensity over time or a comparison of lux levels of pixels including quantification, distribution and size of clusters of pixels meeting certain criteria to a threshold level to make a diagnostic decision, where the light is emitted by the sample and is passed through the emission filter.
  • the present invention is further directed to a related system further comprising software configured to provide processor executable instructions to enable one or more of remote use of the system, an upload of results and other data to a central database or an upgrade of system operating software.
  • the present invention is directed to another related system further comprising one or more excitation filters disposed in front of the one or more light sources.
  • the present invention is directed to yet another related system further comprising a portable, light-tight case having a viewing port disposed in alignment between the emission filter and the camera.
  • the present invention is directed further to a system housed in a housing for detecting the presence of a pathogenic bacteria in a sample in real time.
  • the system comprises a bio-safe container for a sample of interest and a fluorescently-labeled substrate and reagents for an enzyme comprising a pathogenic bacteria that might be present in the sample; said container comprising a mixing mechanism configured to isolate the sample from the substrate/reagents during sample collection prior to mixing the same after collection.
  • the system comprises one or more light sources each having an excitation wavelength of light deliverable to the sample of interest.
  • the system comprises a mobile computing device disposed on the housing such that a camera therein is in alignment with an emission filter positioned between the camera lens and the sample of interest contained in the container where the emission filter is configured to pass fluorescent light emitted by the sample to the camera.
  • the system comprises software configured to provide instructions executable by a processor comprising the mobile computing device to analyze an image of fluorescent light emitted by the sample and captured by the camera via one or both of a comparison of light intensity with a threshold or an increase in intensity over time.
  • the present invention is directed to a related system further comprising one or more excitation filters disposed in front of the one or more light sources.
  • the present invention is directed to another related system further comprising processer executable instructions tangibly stored on the tablet computer to enable one or more of remote use of the system, an upload of results and other data to a central database or an upgrade of system operating software.
  • the present invention is directed further still to a method for detecting a pathogenic bacteria in a sample in real time.
  • obtaining a sample in which the pathogenic bacteria might be present is obtained and is mixed with a substrate specific for an enzyme comprising the pathogenic bacteria in a container, where the substrate is labeled with a reporter that emits fluorescent or colorimetric light upon excitation thereof.
  • An excitation wavelength of light, from one or more light sources, is delivered to the reporter in the sample and light emitted by the reporter is filtered through an emission filter.
  • the filtered emitted light is captured as an image with a camera comprising a mobile device or a mobile computing device and the image is analyzed with software configured to correlate one or more of intensity of light in the image with the presence of the pathogenic bacteria via one or both of a relative comparison to a threshold intensity, shape or distribution of light intensity or a change in intensity over time, thereby detecting the same.
  • the present invention is directed to a related method further comprising the steps of filtering the excitation light with one or more emission filters.
  • the present invention is directed to another related method further comprising one or more steps of enabling remote use of the system, uploading results or other data to a central database or upgrading operating software for the system.
  • the present invention is directed to another related method further comprising diagnosing a pathophysiological condition associated with the pathogenic bacteria.
  • Figures 1A-1 B depict a simple image detection scheme utilizing a mobile device
  • Figures 2A-2B illustrates a custom specimen cup and substrate/reagent cartridge combination with a twist-lock mechanism (Figure 2A) or a stopper mechanism (Figure 2B) for mixing.
  • Figures 3A-3G depict a bio-safe diagnostic device in perspective (Figure 3A), bottom ( Figure 3B) and cross-sectional (Figure 3C) views and illustrate the protocol to prepare a sample for reading ( Figures 3C-3G).
  • Figures 4A-4D depict an alternative bio-safe diagnostic device in unassembled (Figure 4A) and assembled ( Figure 4B) views, and exploded views of the substrate chamber ( Figure 4C) and the read area portion ( Figure 4D) of the device.
  • Figures 5A-5E depicts the results of a CDG-OMe sensitivity test using the detection scheme at control with no substrate (Figure 5A), 2 min (Figure 5B), 10 min (Figure 5C), 20 min (Figure 5D), and 30 min (Figure 5E).
  • Figure 6A-6C show a light tight box (Figure 6A), the box and camera combination with a fluorescent photo before ( Figure 6B) and after lux intensity mapping ( Figure 6C).
  • Figure 7 depicts diagnostic system for detecting bacteria on a surface.
  • Figures 8A-8B shows images of surface samples photographed by an iPhone before (Figure 8A) and after ( Figure 8B) processing via iPhotoLux app.
  • Figure 9 depicts the results from application of a cluster analysis algorithm to samples containing Mycobacteria tuberculosis.
  • the term “a” or “an”, when used in conjunction with the term “comprising” in the claims and/or the specification, may refer to “one”, but it is also consistent with the meaning of "one or more”, “at least one", and “one or more than one”.
  • Some embodiments of the invention may consist of or consist essentially of one or more elements, method steps, and/or methods of the invention. It is contemplated that any device, compound, composition, or method described herein can be implemented with respect to any other device, compound, composition, or method described herein.
  • the term “about” refers to a numeric value, including, for example, whole numbers, fractions, and percentages, whether or not explicitly indicated.
  • the term “about” generally refers to a range of numerical values (e.g., +/- 5-10% of the recited value) that one of ordinary skill in the art would consider equivalent to the recited value (e.g., having the same function or result).
  • the term “about” may include numerical values that are rounded to the nearest significant figure.
  • the terms "computer” or “tablet computer” or “mobile computing device” refer to one or more machines that comprise at least a memory, a processor, a camera or other imaging means, and at least one wired and/or wireless network connection.
  • a computer may comprise a desktop or laptop machine or other electronic media, for example, a smartphone or tablet, as are standard and currently known in the art.
  • any software, modules, applications, add-ons, plug-ins, programs and/or databases, etc. necessary for practice of the invention may be programmed into the computer or tablet computer, may be retrieved over the network connection or may be retrieved from a media storage device tangibly storing the same, may be tangibly stored in computer memory or other electronic media memory and are executable by the processor.
  • a tablet computer are, preferably, a tablet computer for AndroidTM, an operating system by Google Inc. or an iPad ® , a tablet computer by Apple Inc.
  • the term "mobile device” refers to a mobile phone or a smartphone that comprise a camera or other imaging device.
  • the mobile device comprises wireless internet or other network connectivity by which to receive and transmit data or software. This enables capture of the image, transmission and computing analysis external to the mobile device, such as, but not limited to, algorithm analysis in a cloud computing environment.
  • a representative example of a mobile device is, but not limited to, the Apple ® iOS ® -based iPhone ® or Android ® -based devices by Google, Inc.
  • bio-safe refers to the systems, devices and methods provided herein in which a sample is contained in the device sample container such that a health worker, laboratory technician, etc. has no exposure any infectious or contagious agent that may comprise the sample. Once the patient deposits a sample into the sample container and closes the lid, the sample container is not opened nor needs to be opened again to process the sample, as described herein. This comprises a novel feature of the instant invention.
  • a system for detecting the presence of bacteria of interest in a sample in real time comprising means for containing the sample in which the bacteria might be present; means for providing an excitation wavelength of light to the sample; means for capturing light emitted from the sample after excitation; and means for analyzing the captured light.
  • the system may comprise means for housing the system.
  • the means for housing may comprise a portable, light-tight case having a viewing port through which light emitted from the sample after excitation passes.
  • a mobile computing device may be affixed to the light-tight case such that a camera comprising the same is disposed in alignment with the viewing port. Examples of a sample are sputum, saliva, urine, spinal fluid, lymph fluid, pleural fluid, bronchial lavage, stomach lavage, skin, biopsies or a swab of a surface or body orifice
  • the means for bio-safely containing the sample may comprise a sample chamber configured to hold a sample; means for containing a substrate, having a fluorescent reporter, for an enzyme comprising the bacteria and reagents for enzyme activity on the substrate, such that the sample is bio-safely isolated from the substrate/reagents prior to mixing the same; and a mechanism configured to isolate the sample from the substrate/reagents during sample collection prior to mixing and to mix the same after collection, where the containing means comprises a read area after mixing.
  • the sample chamber and the containing means may comprise separate components joinable via the mixing mechanism.
  • the sample chamber and the containing means comprise an integral component having a sample containment portion and a substrate/reagent containment portion separated by the mixing mechanism.
  • the mixing mechanism may comprise a twist, snap, valve, or press-fit mechanism or comprises an interface between the sample chamber and the containing means.
  • the means for providing an excitation wavelength of light may comprise one or more excitation light sources with a wavelength effective to excite a fluorescent reporter comprising a substrate after dequenching thereof for a bacterial enzyme in the sample; a driver for the one or more light sources; and one or more excitation filters disposed in front of the light source.
  • the means for capturing light emitted from the sample may comprise a camera on a mobile device or mobile computing device configured for imaging, processing and connectivity to the internet and an emission filter disposed in line between the sample and the camera; wherein an image of the captured emission wavelength is displayed on a screen thereof.
  • the mobile device or mobile computing device further may comprise software configured to provide processor executable instructions to enable one or more of remote use of the system, an upload of results and other data to a central database or an upgrade or update of system operating software.
  • the camera may capture the emitted light as an image or as pixels.
  • the means for analyzing the captured wavelength may comprise software configured to provide processor executable instructions enabling an assignment of one or more of a colorized scale over a range of lux readings obtained from an image of light emitted from the sample or a comparison of lux levels of pixels including quantification, distribution and size of clusters of pixels meeting certain criteria to a threshold level to make a diagnostic decision.
  • a system for detecting the presence of a pathogenic bacteria in a sample in real time comprising a bio- safe container for a sample, configured to eliminate exposure to infectious bacteria, in which the pathogenic bacteria might be present and a fluorescently-labeled substrate and reagents for an enzyme comprising the pathogenic bacteria; a mixing mechanism configured to isolate the sample from the substrate/reagents during sample collection prior to mixing the same after collection; one or more light sources each having an excitation wavelength of light deliverable to the sample; a camera with camera lens comprising a mobile device or a mobile computing device; an emission filter disposed in alignment between the camera and sample; and software configured to provide instructions executable by a processor to enable analysis of an image of fluorescent light captured by the camera via one or both of a comparison of light intensity with a threshold intensity, shape or distribution of light intensity or an increase in intensity over time or a comparison of lux levels of pixels including quantification, distribution and size of clusters of pixels meeting certain criteria to a threshold level to make a
  • the system comprises one or more excitation filters disposed in front of the one or more light sources.
  • the system comprises a portable, light-tight case having a viewing port disposed in alignment between the emission filter and the camera.
  • the mobile computing device may be affixed to the portable, light-tight case such that a camera comprising the same is disposed in alignment with the viewing port.
  • the system comprises software configured to provide processor executable instructions to enable one or more of remote use of the system, an upload of results and other data to a central database or an upgrade of system operating software.
  • the mixing mechanism may comprise a twist, snap, valve, or press-fit mechanism or may comprise an interface within the sample container.
  • the container may comprise separately a sample chamber and a cartridge containing the substrate/reagents and a mechanism configured to bio- safely join the sample chamber and the cartridge such that the sample is mixed with the substrate/reagents in the cartridge, said cartridge then comprising a read area.
  • the container may comprise a sample containment portion and a substrate/reagents containment portion and a mixing mechanism separating the portions such that upon engaging the mixing mechanism the sample is mixed with the substrate/reagents in the substrate/reagents containment portion, where the substrate/reagents containment portion then comprises a read area.
  • the software may enable instructions to assign a colorized scale over a range of lux readings obtained from the image.
  • the sample may be as described supra.
  • a pathogenic bacteria in a sample in real time comprising the steps of obtaining a sample in which the pathogenic bacteria might be present; mixing the sample with a substrate specific for an enzyme comprising the pathogenic bacteria in a container, said substrate labeled with a reporter that emits fluorescent or colorimetric light upon excitation thereof; delivering, from one or more light sources, an excitation wavelength of light to the reporter in the sample; filtering light emitted by the reporter through an emission filter; capturing the filtered emitted light as an image with a camera comprising a mobile device or a mobile computing device; and analyzing the image with software configured to correlate intensity of light in the image with presence of the pathogenic bacteria via one or both of a relative comparison to a threshold intensity, shape or distribution of light intensity or a change in intensity over time, thereby detecting the same.
  • the method comprises filtering the excitation light with one or more emission filters. Further still to this embodiment the method comprises one or more steps of enabling remote use of the system; uploading results or other data to a central database; or upgrading operating software for the system. Further still the method comprises diagnosing a pathophysiological condition associated with the pathogenic bacteria where light intensity exceeding the threshold or increasing over time correlates to the diagnosis. Examples of a pathophysiological condition are tuberculosis or a staphylococcal infection.
  • the one or more light sources, sample container and emission filter may be positioned within a light-tight housing comprising a viewing port through which the light filtered by the emission filter passes through to the camera.
  • the mobile computing device may be affixed to the light-tight case such that a camera comprising the same is disposed in alignment with the viewing port.
  • the one or more the light source and emission filter may be disposed on the outside of a housing comprising a viewing port through which the light filtered by the emission filter passes, where the sample container is positioned in front of the light source and emission filter.
  • the analyzing step may comprise one or more of assigning a colorized scale over a range of lux readings obtained from the image, an assignment of numerical values with diagnostic thresholds assigned based on pixel lux levels and relative to intensity, shape or distribution of pixels.
  • the sample may be as described supra.
  • the pathogenic bacteria may be a Mycobacteria, Staphylococcus, Escherichia, Salmonella, Shigella, Haemophilus, Pseudomonas, Acinetobacter, Stenotrophmonas, Clostridia, Enterococcus, Legionella, or Listeria.
  • system housed in a housing for detecting the presence of a pathogenic bacteria in a sample in real time, comprising a bio-safe container for a sample of interest and a fluorescently- labeled substrate and reagents for an enzyme comprising a pathogenic bacteria that might be present in the sample; where the container comprises a mixing mechanism configured to isolate the sample from the substrate/reagents during sample collection prior to mixing the same after collection; one or more light sources each having an excitation wavelength of light deliverable to the sample of interest; a mobile computing device disposed on the housing such that a camera therein is in alignment with an emission filter positioned between the camera lens and the sample of interest contained in the container; where the emission filter is configured to pass fluorescent light emitted by the sample to the camera; and software configured to provide instructions executable by a processor comprising the mobile computing device to analyze an image of fluorescent light emitted by the sample and captured by the camera via one or both of a comparison of light intensity with a threshold or an increase in intensity over time.
  • system comprises one or more excitation filters disposed in front of the one or more light sources.
  • system comprises processer executable instructions tangibly stored on the mobile computing device to enable one or more of remote use of the system, an upload of results and other data to a central database or an upgrade of system operating software.
  • the software may provide processer executable instructions to assign a colorized scale over a range of lux readings obtained from the image.
  • the mixing mechanism comprises a twist, snap, valve, or press-fit mechanism or comprises an interface within the sample container.
  • the bio-safe container may comprise separately a sample chamber and a cartridge containing the substrate/reagents and a mechanism configured to bio-safely join the sample chamber and the cartridge such that the sample is mixed with the substrate/reagents in the cartridge, said cartridge then comprising a read area.
  • the bio-safe container may comprise a sample containment portion and a substrate/reagents containment portion and a mixing mechanism separating the portions such that upon engaging the mixing mechanism the sample is mixed with the substrate/reagents in the substrate/reagents containment portion, said substrate/reagents containment portion then comprising a read area.
  • the sample may be as described supra.
  • the diagnostic system comprises means for delivering a specific wavelength or band of wavelengths to a sample in which, if present, an enzyme or bacteria of interest is detected, means for containing the sample with or without a fluorescent substrate, bio-safe means for imaging light emitted by the substrate or the bacteria, and means for analyzing the emitted light.
  • the diagnostic system comprises components for fluorescent or colorimetric spectroscopy and components for imaging and analyzing the fluorescent or colorimetric signal.
  • the diagnostic system utilizes substrates which in the presence of the bacteria or enzymes will fluoresce when an excitation wavelength impinges on the substrate.
  • a small, portable case or box such as a light box, provides a housing for and contains an excitation light source and filters to direct an excitation wavelength at a sample.
  • the emission wavelength emitted by the substrate is imaged by an imaging system or camera integrated into a portable device, for example, but not limited to, a tablet computer which comprises or is affixed to the housing.
  • the emission wavelength emitted by the substrate may be imaged by a mobile device, for example, but not limited to, a camera in a smartphone, with internet connectivity and an app or other software or algorithms for one or more of mapping the lux intensity of the image, an assignment of numerical values with diagnostic thresholds assigned based on pixel lux levels and relative to intensity, shape or distribution of pixels.
  • the diagnostic system also can utilize a colorimetric system. The light source and optics are optimized for colorimetric substrates.
  • the diagnostic system comprises a tablet computer, such as an integrated Android touch screen tablet with a high resolution CCD imaging system, as are standard and known in the art, and a portable, battery backup.
  • the diagnostic system comprises an automobile, i.e., a car charger outlet, and solar battery charger units.
  • the tablet computer is software language customizable and can export data in multiple formats.
  • the diagnostic system comprises a barcode reader that may be used for sample tracking diagnostic device lot number, device lot number threshold adjustments, date code verification and counterfeit device mitigation.
  • the readers are equipped with SIM/LAN capability that allows data to be transferred onto a mobile/computer network.
  • the diagnostic system as configured weighs less than 3 kg and has a small footprint.
  • the diagnostic system can be configured to irradiate a sample within or outside the case comprising the excitation light source.
  • the detection system also may be utilized in the field may be a box similar to the prototype or it may be a miniaturized version of the system that simply snaps on a mobile device, for example, a smartphone, or it may be a custom designed housing with an imaging system, light sources, optics, image capture and app in the custom housing.
  • a tablet computer may be incorporated into the custom designed housing.
  • a light source at the right excitation wavelength is required.
  • the light source may emit at a broader spectrum than desired whereby excitation bandpass filters are utilized to narrow the spectrum of the light source.
  • a light source emitting only in the wavelength required such as a laser or special narrow band LED.
  • a cut off filter that allows all wavelengths above a desired wavelength through to the imaging system may be utilized or a band around the desired wavelength may travel to the imaging system.
  • a fluorescent chip or an optical engine may be utilized instead of the light sources and optics or filters described herein.
  • the fluorescent chip or optical engine may integrate light sources, excitation and emission filtering mechanisms and conversion into a signal into a single chip.
  • an optical engine eliminates the necessity of taking a picture of or imaging the emitted fluorescence light and directly converts it into an electrical signal for analysis.
  • the system can be miniaturized further thereby increasing portability and reducing costs.
  • the portable diagnostic system is designed to detect fluorescence of a fluorescent reporter, e.g., a dye, comprising the substrate specific for the enzyme or associated bacteria of interest.
  • a fluorescent reporter e.g., a dye
  • the fluorescent substrate is brought into contact with the sample in a customized bio-safe diagnostic device.
  • the device may comprise a bio-safe customized specimen cup or sample chamber and substrate/reagent cartridge particularly useful when excitation occurs within the portable case.
  • This diagnostic device provides for mixing of the substrate and sample and interfaces physically with the imaging system.
  • the specimen cup or sample chamber may have an integral configuration with an interface within the chamber to separate the sample and substrate/reagents.
  • the diagnostic devices comprise a bio-safe lock or isolation mechanism, for example, but not limited to, a twist, snap, valve, or press- fit mechanism or a mechanism designed into the device as a bio-safe interface.
  • the substrate may be directly added to a specimen cup containing the sample, for example, when excitation occurs outside the case.
  • a fluorescent imaging system used for identifying bacteria or enzymes on a surface such as, but not limited to, MRSA on operating room equipment or surgical instruments or on a swab utilized to secure a sample from a patient, e.g., from the nostril, may only require a portable LED flashlight with proper excitation filters and the proper emission filter placed over the camera lens.
  • the substrate is sprayed onto the surface or onto the swab and, in a dark room, the substrate will fluoresce.
  • the fluorescence can be viewed through the cell phone viewfinder indicating presence of bacteria or targeted enzyme.
  • the camera function on the phone shows the substrate glowing in the camera monitor.
  • the image may or may not be captured, may or may not require the image analysis and may or may not be transmitted from the hand held device.
  • Imaging the emitted light may be done via a mobile phone or in a mobile device that is designed specifically for this function, i.e., imaging, processing and connectivity to phone system or internet, user interface and diagnostic device interface, that may be similar to a phone but not actually a phone. Either may capture an image or pixel data for analysis. Capturing the image, whether fluorescent or colorimetric, utilizing the high pixel capability of the tablet or mobile device camera enables high sensitivity to be achieved through image processing algorithms that are either customized or apps available off the shelf.
  • the intensity of the imaged light over an established threshold or an increase in intensity of light over time i.e., the slope of increase, or pixel lux level intensity relative to shape or distribution of pixels or all are utilized to detect the presence of bacteria. Detection of bacteria of interest in this manner is indicative of a pathophysiological condition associated with the bacteria.
  • a free off the shelf app named iPhotoLux Soft Energy Consultants Company; www.soft-energy.com
  • the lux mapping software assigns a colorized scale that ranges from the lowest lux reading in the image as magenta and the highest lux reading in the image as white.
  • the sensitivity achieved in the detection system occurs when the range between the lowest lux reading in the image and the highest lux reading is very small and the color scale is assigned over that small range.
  • numerical values may be assigned to the various lux intensity readings that would be run through an algorithm to diagnose the presence or absence of the bacteria or enzyme.
  • An image processing algorithm such as, but not limited to, a "cluster analysis” algorithm, may utilize colorization only, numerical values assigned to pixels with a threshold value, numerical values of the total lux intensity of an area with an overall area threshold, a slope of values over time, pixel lux level intensity relative to shape or distribution of pixels or a combination of any of the above to make a diagnosis of presence or absence of the bacteria or enzyme or for patient data capture.
  • the algorithm affords diagnosis in seconds that is communicated via a colorized image and its utilization is independent of health worker or technician training.
  • the diagnostic system is configured for remote communication and use.
  • the images and results of analysis thereof can be uploaded to a central database.
  • the systems' operating software can be updated.
  • the mobile device or tablet computer comprises an operating system, memory, processor, and is networkable with ubiquitous connectivity.
  • the algorithms or applications comprising the diagnostic system are configured to operate on the mobile device or tablet computer, either can wirelessly port the information, images, analyses or other results to another mobile device or desktop computer by methods well-known in the art.
  • the present invention provides methods for the real-time diagnosis of a pathophysiological condition associated with a pathogenic bacteria or for an in situ determination of the presence of bacterial contamination on a surface or on a patient.
  • the mobility or portability of the diagnostic system enables detection and/or diagnosis in a field environment or within an operating room or any other hospitable setting, for example, the diagnosis of nosocomial infections.
  • Such conditions may be, for example, but not limited to, pneumonia, particularly ventilator-associated or hospital-acquired pneumonia, staphylococcal infections, yeast infections, tuberculosis, urinary tract infections, gastroenteritis, enterococcal infections, or legionnaires' disease.
  • Samples for use in the diagnostic system may be obtained from a biological sample, such as sputum, saliva, urine, spinal fluid, lymph fluid, pleural fluid, bronchial lavage, stomach lavage, skin, or biopsies or from swabbing a surface or body orifice of interest.
  • a biological sample such as sputum, saliva, urine, spinal fluid, lymph fluid, pleural fluid, bronchial lavage, stomach lavage, skin, or biopsies or from swabbing a surface or body orifice of interest.
  • bacteria detectable with the diagnostic system are Mycobacteria, Staphylococcus, particularly methicillin-resistant Staphylococcus aureus (MRSA), Escherichia, Salmonella, Shigella, Haemophilus, Pseudomonas, Acinetobacter, Stenotrophmonas, Clostridia, Enterococcus, particularly vancomycin-resistant Enteroccocus (VRE), Legionella, or Listeria.
  • MRSA methicillin-resistant Staphylococcus aureus
  • Escherichia Salmonella, Shigella, Haemophilus, Pseudomonas, Acinetobacter, Stenotrophmonas, Clostridia, Enterococcus, particularly vancomycin-resistant Enteroccocus (VRE), Legionella, or Listeria.
  • FIG. 1A is a cartoon illustration of one embodiment of the diagnostic detection system.
  • a case 110 such as a light box case comprises a LED light source 120 with an LED driver.
  • the LED light source has an excitation bandpass filter 125 to filter the LED light to pass only the required excitation wavelength 130.
  • An emission filter 140 is disposed proximate to a viewing port 115 on a surface 112 of the case and a handheld device 150, such as a mobile phone is disposed on the outside of the surface 112, such that the camera 155 in the phone aligns with the emission filter.
  • the case can receive a container 160 containing a sample 165, mixed with a fluorescently labeled substrate for an enzyme associated with a bacteria of interest that might comprise the sample.
  • the container with sample is positioned within the case to both receive the excitation light from the LED and to emit fluorescent light 135 which is filtered through the emission filter and captured by the phone camera. Analysis of the captured image is performed with software to colorize the image based on Lux intensity relative to a calibrator.
  • a cyan LED with a 1 15v driver is filtered through a 490 nm excitation filter with a 20 nm bandpass.
  • the filtered excitation wavelength impinges on a sample in which the bacterial enzyme beta-lactamase (BlaC) is detected.
  • the sample comprises a BlaC substrate modified with a fluorescent dye. Interaction of the substrate with BlaC un-quenches the dye resulting in fluorescent light emission when excited.
  • the emitted light is filtered with a 530 nm emission filter with a 43 nm bandpass whereupon the image is captured by the camera on a mobile phone.
  • Subsequent analysis with software can, for example, colorize the image based on Lux intensity relative to a calibrator and the colorized image can be viewed on the phone's screen and/or wirelessly transmitted to another mobile device or computer for analysis, storage and/or review.
  • Figure 1 B is a cartoon illustration of a preferred embodiment of the diagnostic detection system.
  • the case 110 comprises the LED light source 120 with an LED driver and the excitation bandpass filter 125 to filter the LED light to pass only the required excitation wavelength 130.
  • a tablet computer 170 comprising a camera aperture or other imaging aperture 175 is disposed or affixed to the outside of surface 112 on the case and replaces the viewing port 115 (see FIG. 1A).
  • the emission filter 140 is disposed proximate to and in alignment with the aperture 175.
  • the case 110 can receive at 180 the bio-safe diagnostic device 160 with sample 161 and the fluorescently labeled substrate 163 for an enzyme associated with the bacteria of interest that might comprise the sample.
  • the diagnostic device with sample and enzyme is positioned to receive the excitation light 130 from the LED after the enzyme is mixed with the sample to form the mixed sample 165.
  • the emitted fluorescent light 135 which is reflected by mirrors 190a,b, disposed on inner surface 114, to the emission filter 140, is filtered through the emission filter and into the aperture 171 and is captured by the camera or imaging device comprising the tablet computer.
  • the captured image is analyzed, stored and/or transmitted, as described herein. It is to be noted that any combination of the excitation light 130 from the LED after the enzyme is mixed with the sample to form the mixed sample 165.
  • the emitted fluorescent light 135 which is reflected by mirrors 190a,b, disposed on inner surface 114, to the emission filter 140, is filtered through the emission filter and into the aperture 171 and is captured by the camera or imaging device comprising the tablet computer.
  • the captured image is analyzed, stored and/or transmitted, as described herein. It is to be noted that any combination of the excitation light 130 from
  • Figures 2A-2B depict customized bio-safe diagnostic devices for containing a sample and a fluorescently labeled substrate and the method of its use.
  • the diagnostic device comprises a customized specimen or sample cup 210 and a customized substrate/reagent cartridge 220.
  • the specimen cup and cartridge each have a surface 210a, 220a that can be mated via a twist-lock attachment mechanism or may be one integral piece with a valve mechanism to allow the sample to flow to the read area of the cartridge where the fluorescently labeled substrate is contained.
  • the specimen cup has a bio-safe design. Once a sample, e.g., a biological sample, such as sputum, is placed within the bio-safe specimen cup, it never leaves the cup during the entire detection procedure.
  • sputum or other sample type is placed in the specimen cup at 230 and the required substrate and any necessary reagents are placed in the cartridge at 235.
  • the specimen cup and the cartridge are twist-locked together or the valve is opened with the integral design, either allowing the sample to flow into the read area, agitated for mixing of sample, reagents and substrate allowed to sit for about 10 mins at 240. This enables batching of tests or, alternatively, two technicians can work together to perform many tests per hour.
  • the mixed sample and substrate in the specimen cup/cartridge are placed within the light box case 110 at 250 whereupon the excitation light is directed to sample/substrate mixture in the read area and emitted light is transmitted out of the read area to a mobile camera or other device, as described in more detail in Figures 1A-1 B.
  • a result is obtained and viewed on the screen of a mobile phone or tablet computer.
  • the bio-safe diagnostic device may comprise a single container 260 with cap 265 which comprises a sample containment portion 262 and a substrate containment portion 264 into which the fluorescently labeled substrate 280 and any reagents required for enzyme activity when mixed with the sample 285 are pre-loaded.
  • the containment portions are separated by, in combination, a divider 270 comprising a material that dimples under pressure and a stopper 275 having an upper component 276 that aligns with the divider and a lower component 278 that dimples the divider material. This combination of the divider and the stopper prior to mixing isolates the sample from the substrate.
  • sputum or other sample type 285 is placed in the sample containment portion of the container and is isolated from the substrate by the downwardly positioned stopper. Upward pressure at 290 pushes the dimple and stopper into the sample containment portion, thereby allowing mixing of the sample and substrate within the containment portion. At this point in the process the containment portion functions as a read area 295, as described in Figure 2A.
  • Figures 3A-3G depict a preferred bio-safe diagnostic device for use in the diagnostic detection system.
  • the device 300 comprises a container 310, a removable lid 320 disposed in covering relationship to the container and a movable read area cartridge 330 disposed proximate to the container.
  • the container comprises a plurality of finger grips 312a,b,c,d,e,f disposed uniformly around the outside surface thereof.
  • the removable lid has a recessed upper surface portion 321 comprising an area 322 upon which patient information, such as a patient identification number, can be written.
  • the lid further comprises a lower portion 323 for securely engaging the open upper end of the container.
  • the lid has an outer diameter d1 across the upper surface portion of about 70 mm and an outer diameter d2 across the lower portion of about 53 mm.
  • the height h of the device with the lid in a closed configuration is about 44 mm.
  • the read area cartridge 330 is protected by a removable read area cover 340 disposed in a protective covering relationship around the read area.
  • Figure 3B is a bottom view of the device 300 without the read area cover 340.
  • the bottom surface of the read area cartridge 330 comprises a bar code 332 which can be scanned by a barcode reader comprising the diagnostic device for sample tracking.
  • the finger grips 312a,b,c,d,e,f are more readily viewable.
  • Figure 3C is a longitudinal cross-sectional view of the device 300.
  • the lid 320 is removed from the container 310.
  • This cross-sectional view depicts the means 324 for securely engaging the open end of the container 310 and the corresponding receiving means 314 by which to secure the lid.
  • the lid further comprises a compartment 325 depending from the upper recessed surface portion 321 having a pierce seal 326, for example a foil, disposed across the opening at the lower end of the compartment to contain the TSS wet reagents 328 therein and a means 327 for metering a sample disposed around the outer surface proximate to the seal.
  • a pierce seal 326 for example a foil
  • the container 310 has a lower portion 316 substantially funnel shaped in which to receive a sample, for example, a sputum sample, from a patient.
  • the lower or stem end of the funnel portion of the container has a pierceable seal 317, for example, a foil seal, disposed across the opening to contain the sample and, subsequently, the sample mixed with the wet reagents.
  • the funnel shaped portion has dimensions sufficient to securely receive the lower end of the wet reagents compartment therein.
  • the inner surface of the stem end comprises means 318 for receiving and securing the movable read area component thereto.
  • the movable read area cartridge comprises TSS lyophilized reagents 333 and REFS lyophilized reagents 334 and is adapted to receive the sample-wet reagents mixture.
  • the movable read area cartridge 330 comprises corresponding engagement means 338 to movably engage the receiving means 318 to raise the cartridge.
  • Figures 3D-3G illustrate the protocol for preparing a sample.
  • a sputum sample is placed into the container portion 316.
  • Figure 3D illustrates the second step in which the lid 320 is closed with engagement/receiving means 314, 324 securely engaged.
  • the wet reagents compartment 325 is thereby in contact with the sample such that the desired quantity is metered automatically via the metering means 327 at 350a, b into the stem end of lower portion 316, but separated from the read area by seal 317.
  • step 3 as illustrated in Figure 3E, closing the lid 320 concomitantly pierces the foil seal 326 which allows the TSS wet reagents 328 to mix with the metered sample at 360 thereby yielding a treated sample. At this point the sample may be transported, if necessary, for reading.
  • step 4 as illustrated in Figure 3F, the read area cartridge 330 is engaged with the lower portion 316 of the container 310 which pierces the foil seal 317 so that the treated sample mixes with the TSS 333 and REFS 334 lyophilized reagents at 370.
  • step 5 as illustrated in Figure 3G, the read area cover 340 is removed and the device 300 with prepared sample at 370 is inserted into a diagnostic detection device, as described herein.
  • FIG. 4A depicts an alternative bio-safe diagnostic device.
  • the bio- safe diagnostic device 400 comprises a container 410 having a reagent chamber 415 with bio-seal 416 and a read area portion 420, a sample collection wand 430, having a threaded bio-safe cap 431 and a sample containment end 432, and a disposable read area cover 440.
  • the assembled bio-safe diagnostic device has a length L of about 5.5 cm to about 14 cm.
  • the container 410 has a label area 411 on which patient information can be written and means 412 for bio-safely receiving the bio-safe cap comprising corresponding means 433 for engaging the container.
  • the container comprises all the reagents necessary for diagnosis or detection of a pathogenic bacteria in a sample in the reagent chamber.
  • the wet reagents and lyophilized reagents are separated by seals, such as foil seals, which are pierceable when a sample is to be read (see Figure 4C).
  • the container is light tight, can be stored on the shelf and packaged in bulk.
  • Figure 4B depicts the sample collection wand 430 disposed inside the container 410 and securely and bio- safely engaged with the receiving means of thereof.
  • the read area cover 440 is removed to view the read area portion 420.
  • the bio-safe sample collection wand 430 comprises a plurality of grips, represented at 434 disposed around the outer surface.
  • a reagent shield 435 is disposed proximate to the sample containment end 432 such that when the wand is placed into the container 410, the reagent shield is disposed to contain the sample during mixing with the reagents.
  • the sample containment end comprises a grooved collection area 436 which can collect about 500 microliters of sample, for example, sputum. The grooves capture a thick sample mechanically and can capture a thin sample with capillary action.
  • the bio- safe sample collection wand can be packaged in bulk and stored on a shelf because it contains no reagents.
  • the bio-safe diagnostic device is low cost and easy to use with minimal training.
  • the bio-safe device comprises no metal or glass and is constructed of injection molded plastics.
  • the bio-safe diagnostic device is designed for open bench use and is suitable for incineration.
  • FIG. 4C depicts the components of the reagent chamber 415 in cross-section.
  • the reagent chamber comprises a liquid reagent portion 417, a lyophilized reagent portion 418. Seals 419a, b on both ends of the liquid reagent portion separate it from the rest of the container and from the lyophilized reagent portion. Seals 419b, c separate the lyophilized reagent portion liquid reagent portion from the read area portion 430.
  • the reagents comprising the reagent chamber can be stored in the container 410 at 35 °C for about 24 months. Seals 419a, b are pierceable by the wand 420 and seal 419c is pierceable by the read area tip (see Figure 4D). This enables the sample and reagent system to mix.
  • Figure 4D depicts the components of the read area portion 420 of the container 410.
  • the read area portion comprises a read tip 421 having a bar code 421 a disposed thereon and a read area 422.
  • the bar code may contain lot specific data and may be utilized for device specific tracking and/or anti-counterfeit verification.
  • a seal piercing tip 426 is disposed over the cellulose strip at 423a.
  • the cellulose strip with seal piercing tip is disposed within the read tip such that the read area 423b is contained within the corresponding read area 422 in the read tip 421 and the seal piercing tip is disposed proximate to the seal 419c in the container 410.
  • a seal 427 is disposed over the read tip to contain the sample + reagents + substrate mixture when seals 419a,b,c are pierced.
  • the assembled read area portion is covered with disposable cover 440.
  • a user writes patient information on the label 411 , collects a raw sample with the sample containment end of the wand 430 and inserts the wand into the container 410. This puts the reagent shield 435 in place prior to mixing with the reagents.
  • the user seats the bio- safe cap 431 by engaging the engaging means 433 with the corresponding receiving means 412 in the container. Seating the wand pierces the seals 419a, b such that the sample mixes with the liquid reagents in the reagent portion 417 and the lyophilized reagents in the reagent area 418.
  • Queue time in the reagent chamber may be from less than 20 minutes to hours or days.
  • the read area portion 420 of the container is unlocked and activated.
  • the seal piercing tip 426 pierces seal 419c whereupon the sample + reagent mixture flows into the read area portion and contacts any other necessary reagents 424 and the substrate 425 on the cellulose strip 423.
  • the read tip cover 440 may be removed and the bio-safe diagnostic device with sample may be inserted into the diagnostic system for reading as described herein.
  • Figures 5A-5E depict the results of a CDG-OMe sensitivity test.
  • BCG Bacille Calmett-Guerin
  • Figure 5A shows BCG dilutions in sputum with no substrate.
  • BCG beta-lactamase is detected with the CDG-OMe substrate at 2 mins, 10 mins, 20 mins, and 30 mins (Figs. 5B-5E) after addition of the CDG-OMe using the detection system described herein with an excitation range of about 480-500 nm and an emission range of about 509 to 552 nm.
  • the limit of detection is about 10 CFU.
  • the images were obtained with an iPhone 4S and analyzed with the off the shelf app. A blue color indicates the absence of the enzyme, as shown in the samples with no or 0 bacteria. An increase in fluorescence, correlating to an increase in bacteria, is demonstrated by an increase in orange color. Thus, by using a reference with 0 bacteria and setting the threshold it is demonstrated that dilutions with ⁇ 10 bacteria and higher are visibly detectable using the light intensity algorithm. This enables a diagnosis of the pathophysiological condition associated with the detected bacteria.
  • Figures 6A-6C depict the physical layout of a detection system with or without images displayed on a mobile phone screen.
  • Figure 6A shows a light tight box with mounting brackets that positions a mobile phone camera over the viewing port through which light emitted from the sample after excitation is transmitted.
  • the viewing port comprising the emission filters is disposed such that when a mobile phone is slotted into the brackets, the camera is aligned with the viewing port to image the light emitted from the sample.
  • Figure 6B shows a photo of a visible fluorescent image initially obtained by the camera and
  • Figure 6C shows the Image after lux intensity mapping utilizing an off-the shelf iPhone app (iPhotoLux)
  • Figure 7 depicts the physical layout of a detection system where the LED light source and filter systems are configured to detect bacteria or associated enzymes in samples outside the light box case.
  • the detection system utilize two LED light sources with the excitation filters mounted in front of the LEDs.
  • the mobile device camera is mounted on the outside of the box behind the single emission filter.
  • the samples are positioned in front of the LEDs and emission filter to receive the excitation light from the LEDs and to transmit any emitted light through the emission filter to the camera for imaging.
  • LED selection is based on optimizing the emission of the excitation wavelength, centered at 505 nm.
  • Two LEDs Philips Lumileds Lighting Company, model number LXML-PE01-0070 Cyan Rebel LEDs pre-soldered to two Saber 20mm star MCPCB bases
  • the two excitation optical lens utilized was were 25.4 mm diameter bandpass filters with 490 ⁇ 10 nm centers and 10 ⁇ 2 nm FWHM.
  • the maximum excitation wavelength reaching the sample, including tolerance, is 504 nm. Selection of this bandpass range and tolerance minimizes any excitation of the sample below 476 nm to minimize any non-substrate associated fluorescence emitted from the sample.
  • Utilizing this LED and optics system to optimize the excitation wavelength and to filter the emission wavelength enables image capture with high sensitivity being achieved that is specific to the targeted excitation and emission spectrum of the specific substrate.
  • Different substrates may utilize the same or different combinations of excitation light sources, excitation filters and emissions filters to achieve the desired result.
  • Figures 8A-8B show the unprocessed fluorescent image and the image analyzed using iPhotoLux.
  • An actual TB negative sample was used to set the baseline lux intensity threshold.
  • a baseline lux intensity threshold is determined and utilized to perform diagnosis or to the indicate presence of the bacteria or target enzyme. For example, if through experimentation the lux intensity of an image of a sample mixed with substrate free of bacteria or enzyme is set, and that minimum lux intensity level is seen as, e.g., blue in the image, then any sample generating an image that upon running through the image processing algorithm indicates as yellow, orange or white, indicates the presence of the bacteria or enzyme. A sample mixed with substrate after image processing that shows as magenta or blue or possibly green would be diagnosed as free of the bacteria or enzyme.
  • Figure 9 illustrates a cluster analysis of images at lux levels of pixels for a negative control (left), less than 10 colony forming units (middle) and 10-100 colony forming units (right). This includes quantification, distribution and size of clusters of pixels meeting certain criteria that are then compared to a threshold level to make a diagnostic decision. Particularly, the algorithm uses multiple characteristics including pixel LUX intensity, shape and size of fluorescent clusters and automatically assigns a value. Diagnosis is achieved in seconds and is communicated with colorized images.

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Abstract

La présente invention concerne un système de diagnostic portable qui comporte un boîtier opaque qui présente un regard et qui contient un récipient d'échantillon biologiquement sûr, une source de lumière d'excitation ayant un filtre, un filtre d'émission et un dispositif mobile ou un dispositif informatique mobile ayant une caméra. La lumière d'excitation excite un substrat marqué par la fluorescence propre à une bactérie d'intérêt qui peut comporter l'échantillon après avoir désactivé celui-ci. La lumière fluorescente émise est filtrée par le filtre d'émission et capturée en tant qu'image par la caméra. L'analyse, par exemple l'analyse de lux, est faite par une application ou par un autre logiciel équivalent, contenu dans le dispositif mobile ou dans le dispositif informatique mobile. L'apparition de couleur est en corrélation avec la présence des bactéries, ce qui est considéré comme étant un diagnostic d'un état pathophysiologique associé à celles-ci.
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CN106404733A (zh) * 2016-09-18 2017-02-15 天津博硕东创科技发展有限公司 基于移动智能终端的食品安全检测装置及检测方法
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CN110830768A (zh) * 2019-10-31 2020-02-21 北京君立康生物科技有限公司 一种菌落图像的采集处理方法及装置
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US20210349033A1 (en) * 2020-05-07 2021-11-11 Thomas James Riha Intelligent reader and data recording system for biological, chemical, and instant biological and enzymatic sterilization indicators
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Cited By (12)

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Publication number Priority date Publication date Assignee Title
JP2015205841A (ja) * 2014-04-22 2015-11-19 住友化学株式会社 キサンテン化合物及びその用途
CN106404733A (zh) * 2016-09-18 2017-02-15 天津博硕东创科技发展有限公司 基于移动智能终端的食品安全检测装置及检测方法
CN106404733B (zh) * 2016-09-18 2023-08-22 天津博硕东创科技发展有限公司 基于移动智能终端的食品安全检测装置及检测方法
CN111630604A (zh) * 2017-11-20 2020-09-04 纳诺全球公司 基于生物细胞或生物物质的检测的数据收集和分析
WO2020033593A1 (fr) 2018-08-07 2020-02-13 Britescan, Llc Dispositif de balayage portable pour déterminer des attributs de matériaux d'échantillon
US10684231B2 (en) * 2018-08-07 2020-06-16 Britescan, Llc Portable scanning device for ascertaining attributes of sample materials
US11243172B2 (en) * 2018-08-07 2022-02-08 Britescan, Llc Portable scanning device for ascertaining attributes of sample materials
EP3833242A4 (fr) * 2018-08-07 2022-08-03 Britescan, LLC Dispositif de balayage portable pour déterminer des attributs de matériaux d'échantillon
CN110830768A (zh) * 2019-10-31 2020-02-21 北京君立康生物科技有限公司 一种菌落图像的采集处理方法及装置
CN110830768B (zh) * 2019-10-31 2021-07-16 北京君立康生物科技有限公司 一种菌落图像的采集处理方法及装置
US20210349033A1 (en) * 2020-05-07 2021-11-11 Thomas James Riha Intelligent reader and data recording system for biological, chemical, and instant biological and enzymatic sterilization indicators
US12007332B2 (en) 2022-01-21 2024-06-11 Britescan, Inc. Portable scanning device for ascertaining attributes of sample materials

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