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WO2009152209A2 - Test de détection d'analyte visuel/par fluorescence combiné - Google Patents

Test de détection d'analyte visuel/par fluorescence combiné Download PDF

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Publication number
WO2009152209A2
WO2009152209A2 PCT/US2009/046848 US2009046848W WO2009152209A2 WO 2009152209 A2 WO2009152209 A2 WO 2009152209A2 US 2009046848 W US2009046848 W US 2009046848W WO 2009152209 A2 WO2009152209 A2 WO 2009152209A2
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WO
WIPO (PCT)
Prior art keywords
fluorescing
sample
visible
test
target
Prior art date
Application number
PCT/US2009/046848
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English (en)
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WO2009152209A3 (fr
Inventor
Uma Mahesh Babu
Robert W. Vandine
Robert P. Sambursky
Ganga Kanaujia
Richard Rivas, Jr.
Original Assignee
Rapid Pathogen Screening, 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 Rapid Pathogen Screening, Inc. filed Critical Rapid Pathogen Screening, Inc.
Publication of WO2009152209A2 publication Critical patent/WO2009152209A2/fr
Publication of WO2009152209A3 publication Critical patent/WO2009152209A3/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
    • G01N33/543—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

  • the invention pertains to the field of lateral flow immunoassays. More particularly, the invention pertains to lateral flow immunoassays with increased sensitivity due to the use of fluorescent markers.
  • Lateral flow immunoassays are a subset of antibody/antigen-based immunoassays combining various reagents and process steps in one assay strip, thus providing a sensitive and rapid means for the detection of target molecules.
  • Lateral flow immunoassays are available for a wide area of target analytes and can be designed using sandwich or competitive test principles. Generally high molecular weight analytes with several epitopes are typically analyzed in a sandwich format whereas small molecules representing only one epitope are usually detected by means of a competitive assay. The first tests were made for human chorionic gonadotropin (hCG). Today there are commercially available tests for monitoring ovulation, detecting infectious disease organisms, analyzing drugs of abuse and measuring other analytes important to human physiology. Products have also been introduced for veterinary testing, environmental testing and product monitoring.
  • hCG human chorionic gonadotropin
  • United States Patent 5,714,341 discloses a lateral flow immunoassay for HIV specific antibodies in saliva samples.
  • the saliva sample is diluted in a sample buffer and the lateral flow immunoassay is dipped into the diluted saliva sample.
  • the present invention enhances the sensitivity of visually read lateral flow immunoassay tests by adding a small quantity of a fluorescing element, such as a fluorescing dye or fluorescing latex bead conjugates, to the initial conjugate material.
  • a fluorescing element such as a fluorescing dye or fluorescing latex bead conjugates
  • the visible spectrum test line is visible to the unaided eye, the test result is observed and recorded.
  • a light of an appropriate spectrum such as a UV spectrum
  • the UV light will excite and fluoresce the fluorescing dye or the fluorescing latex beads which are bound in the test line to enhance the visible color at the test line. If the test is negative, there will be no fluorescing elements to excite, and there will therefore be no enhancement.
  • Figure 1 shows a sample analysis device in an embodiment of the present invention.
  • Figure 2 shows a housing containing the strip of Figure 1.
  • Figure 3 shows a collection device for collecting a sample.
  • Figure 4 shows a test kit including the sample analysis device of Figures 1 and 2 and the collection device of Figure 3.
  • Figure 5 shows another embodiment of a sample analysis device of the present invention.
  • Figure 6 shows examples of test results and how using fluorescence enhances the ability to interpret the test result for weak positives.
  • Figure 7 shows a close up view of a positive fluorescent test result for the weakly positive visual test shown in Figure 6.
  • Figure 8 shows a close up view of another positive fluorescent test result for a weakly positive visual test.
  • the present invention enhances the sensitivity of visually read lateral flow immunoassay tests by adding a small quantity of at least one fluorescing element, including, but not limited to, a fluorescing dye or fluorescing latex bead conjugates, to the initial conjugate material.
  • a fluorescing element including, but not limited to, a fluorescing dye or fluorescing latex bead conjugates
  • a fluorescent element can be "masked” by attaching the appropriate "quencher” moiety in the appropriate vicinity.
  • the fluorescence is "unmasked”. This phenomenon is called Fluorescence Resonance Energy Transfer (FRET).
  • FRET Fluorescence Resonance Energy Transfer
  • the methods and devices discussed herein use both a visual label and a fluorescent label together in the same assay in order to detect a target molecule.
  • the fluorescent element is preferably used as a real time supplement to the visual indicator in a rapid result point of care sample analysis device.
  • the fluorescent labels preferably increase the sensitivity of the test at least tenfold, without sacrificing the rapidity and simplicity of the test.
  • the analytical tests discussed herein preferably permit a result while the patient is still being examined by the practitioner.
  • the test result is obtained in under 10 minutes after applying the sample to the device, and it is preferably read at approximately 10 minutes. In samples that are highly positive, a readout of the test zone (preferably a test line) is visible within approximately 5 minutes.
  • the visible labels and the fluorescing elements are each coupled with at least one specific binding partner for the target molecule in the biological sample.
  • the specific binding partners for the analytes in the sample are monoclonal, polyclonal or recombinant antibodies or fragments of antibodies capable of binding to a pathogen.
  • the specific binding partners may also be antigens capable of binding to antibodies against a pathogen or an allergen.
  • Other types of binding partners include, but are not limited to, bioorganic macromolecules like aptamers or receptors, nanoparticles, or nucleic acids.
  • Preferred targets include, but are not limited to, proteins, glycoproteins, proteoglycans, and lipoproteins.
  • Other preferred targets include, but are not limited to, pathogens, low-molecular- weight compounds, and/or allergy-associated components.
  • the pathogens are preferably selected from viruses, microorganisms, e.g. bacteria and parasites, e.g. amoebae or nematodes.
  • the allergy-associated components are preferably selected from allergens and anti-allergic components.
  • the target is on the surface of the cell or organism, so it is not necessary to lyse the cells to detect the target.
  • the sample is preferably a sample of body fluid.
  • the sample of body fluid is preferably taken from a body surface selected from mucosal membrane fluids (preferably of the oral, nasal, vaginal, and ocular cavities), blood, urine, tears, secretions from glands and secretions from lesions or blisters, e.g. lesions or blisters on the skin. More preferably, the sample is selected from oral, nasal, ocular, genital and rectal fluid, secretions from skin lesions or blisters, CFD (cerebral spinal fluid), and exudates.
  • mucosal membrane fluids preferably of the oral, nasal, vaginal, and ocular cavities
  • the sample is selected from oral, nasal, ocular, genital and rectal fluid, secretions from skin lesions or blisters, CFD (cerebral spinal fluid), and exudates.
  • U.S. Published Patent Application No. 2005/0175992 discloses a method for detecting targets, such as pathogens and/or allergy-associated components, in a human body fluid where the body fluid sample is collected by a collection device, such as a swab member. The samples are transferred from the swab member to a sample analysis device, on which an analysis of the targets can occur by immunochemical or enzymatic means. The test result is capable of being displayed within a very short period of time and can be directly read out by the user.
  • the inventions disclosed in this copending application are particularly advantageous for the diagnosis of conjunctivitis.
  • the chromatographic test strip shown in Figures 1 through 4 includes a plurality of different strip materials.
  • the device preferably includes an absorbent pad (1), an application zone (2), a detection zone (3) and a waste zone (4).
  • the strip materials are arranged on an adhesive plastic backing (5).
  • the absorbent pad (1) is provided in this example for adding an elution medium in order to facilitate the transfer of the sample to the detection zone (3).
  • US Patent Publication No. 2007/0059682 describes methods to increase specificity of lateral flow immunoassays. These methods could also be used in combination with the embodiments described herein.
  • Figure 2 shows a housing (6), which is preferably plastic, containing the strip as shown in Figure 1.
  • a sample application window (7) brings a collection device into contact with the strip.
  • the test result is displayed in the read out window (8).
  • Figure 3 shows the collection device for collecting a sample.
  • the collection device is a swab member.
  • the collection device includes a body (9), which is preferably plastic, with a sample collection material (11) fixed on it and an opening (10) corresponding to a read out window when the collection device is operatively in contact with a test strip.
  • Figure 4 shows a test kit, which includes the sample analysis device of Figures 1 and 2 and the collection device of Figure 3.
  • the chromatography test strip (100) includes an application zone (or sample zone) (101). The sample is applied to the application zone (101).
  • the test strip also includes a detection zone (105).
  • the detection zone (105) includes at least one test zone (102), which is preferably a test line. Although only one test line is shown in the figure, multiple test lines are within the spirit of the invention. In some embodiments where there are multiple targets, the presence of each target preferably corresponds to a separate test line. In other embodiments where there are multiple targets, the presence of multiple targets may be indicated on the same test line such that the presence of more than one target has different characteristics than the presence of a single target. For example, the presence of multiple targets on the same test line may be visually indicated by a different color than the presence of each of the targets alone.
  • the test strip (100) also preferably includes at least one control zone (104), which is preferably a control line. As shown in Figure 5, the control zone (104) is preferably downstream of the test zone (102). However, in other embodiments, the control zone (104) may be located upstream of the test zone (102).
  • a fluorescing element is coupled to the specific binding partner for the target or targets of interest.
  • the visual label is also coupled to the specific binding partner for the target or targets of interest.
  • the specific binding partner is an antibody or antigen.
  • the specific binding partner includes a plurality of nanoparticles or a nucleic acid.
  • the fluorescing element coupled to the antibody or antigen as well as the general blocking agent, for example Bovine Serum Albumin (BSA), is added to the antibody or antigen coupled to the visual label.
  • BSA Bovine Serum Albumin
  • the visual label may be any label visible to the naked eye, including, but not limited to, colored particles such as colloidal gold, dyed latex beads, selenium, or carbon.
  • the nanoparticles that may be used include, but are not limited to, selenium, carbon, and colloidal gold.
  • the visual tags are also coated with fluorescing elements.
  • the fluorescing element is a fluorescing dye.
  • a mixture of preferably colorless fluorescing latex bead conjugates are mixed with colloidal gold (a visible spectrum) conjugates, or conjugates producing a visible read test line, in lateral flow immunoassays to enhance sensitivity of the assay and to aid in visually reading true positives and true negatives.
  • visually red particles such as colloidal gold or red dyed latex beads
  • the antibody or antigen is also coupled to a fluorescing dye.
  • the conjugated or coupled antibody or antigens are blocked with a mixture of a general blocking agent and a fluorescing dye blocking agent.
  • the blocking agent is bovine serum albumin.
  • red colored colloidal gold or red dyed latex bead conjugates and colorless fluorescent latex beads that fluoresce to produce a red color under UV excitation are used.
  • the red colored colloidal gold or red dyed latex bead conjugates are conjugated with the antibody or antigen specific to the target and the antibody or antigen is also coupled to the colorless fluorescent latex beads.
  • the conjugated or coupled antibody or antigens are blocked with a mixture of a general blocking agent and a fluorescent latex bead blocking agent.
  • the blocking agent is bovine serum albumin.
  • any color may be used for the dyed latex beads, the fluorescing dye, and/or the fluorescing beads.
  • the same color does not necessarily need to be maintained, so that the fluorescing dye or the fluorescing beads when excited can have a different color than the gold labeled particles or the dyed latex beads.
  • Other colorless fluorescent beads which can be excited at other wavelengths including those in the visible spectrum and those that will fluoresce to emit colors of the visible spectrum other than the red color mentioned above can also be used in the present inventions to achieve greater sensitivity with the fluorescent beads.
  • One of the preferred embodiments is made by coupling the specific binding partner, for example an antibody or antigen, mixed with fluorescing dye and then conjugating the coupled antibody or antigen to the colloidal gold or red dyed latex beads.
  • Another preferred embodiment is made by adding 1% by volume of colorless UV fluorescing latex bead conjugates that fluoresce to produce a red color to the optimum quantity of the colloidal gold (visible spectrum) conjugates.
  • the 1% colorless fluorescing latex beads enable visual reading of the test line with a minimum of a tenfold increase in sensitivity at the lower limit of detection of the visible spectrum labeled conjugate.
  • the UV fluorescing beads fluoresce with varying luminosity depending on the wavelength of the excitation source. A greenish lower luminosity is excited with 395 nm light. The "gold" colored band with a higher luminosity is excited with 365 nm light. It is well within the skill of those in the art to choose a color and a light source to achieve a test result readily visible under a fluorescent light source.
  • the fluorescing latex beads are preferably colorless under ambient light so as to not interfere with the reading of a strong positive result. However, it will be readily appreciated that the fluorescing latex beads can alternatively be lightly colored.
  • the fluorescing latex beads typically have a diameter of 1 to 500 nm, preferably 20 to 400 nm.
  • fluorescing latex beads or the fluorescing dye preferably fluoresce at ultraviolet light wavelengths (IOnm to 400nm)
  • any system where the beads or the dye will be excited and fluoresce at other wavelengths could also be utilized in the present invention.
  • wavelengths in the range of 480 nm to 490 nm are used.
  • Alternative wavelengths within the spirit of the present invention include, but are not limited to, all visible (400nm to 700 nm) and infrared (750 nm and 100 ⁇ m) spectra.
  • the fluorescing latex beads should be mobile to be transported along the test strip until captured in the test zone and/or the control zone.
  • Suitable examples of beads that can be used in the present invention include carboxylated latex beads, latex beads that can couple to proteins passively and beads that can be coupled to proteins by other chemistries.
  • the fluorescing latex beads are preferably mobile on the immunoassay test strip under the action of a suitable buffer, which permits the use of small volumes of analyte, but in the case of a body fluid such as urine, the body fluid itself may cause transport of the fluorescing latex bead conjugates, as well as the gold label conjugates, along the test strip.
  • the visible luminosity of the visually read test line (102) is determined by the number of gold label particles that accumulate at the test line.
  • the sensitivity of the visually read lateral flow immunoassay test is enhanced by adding a small quantity of the fluorescing element, for example a fluorescing dye or fluorescing bead conjugates, to the initial conjugate material (such as gold labeled conjugate material or dyed latex bead conjugate material) and can be excited with a fluorescent light source, for example a small UV light, to provide an aid in clarifying a true positive versus a true negative test result.
  • the fluorescing element for example a fluorescing dye or fluorescing bead conjugates
  • the fluorescing latex beads which are used in the present invention are preferably colorless in the visible spectrum, and fluoresce under UV light. These latex beads are conjugated to an antibody or other binding partner which reacts with a capture analyte at the test line (102) in the same manner as the gold labeled particles, so that both the fluorescing latex beads and the gold labeled particles accumulate at the test line (102).
  • a suitable amount of the UV fluorescing latex bead conjugates is preferably 0.1% to 10% by volume, based on the volume of the visible colloidal gold or dyed latex bead conjugates, and preferably is about 1% by volume of the visible colloidal gold or dyed latex bead conjugates.
  • the visible spectrum test line (102) is visibly present, the test result is observed and recorded, and no ultraviolet light excitation is required.
  • a UV spectrum light is cast on the test line to excite and fluoresce the fluorescent latex beads to indicate the presence of the fluorescent latex beads bound in the test line (102).
  • the limit of detection for a visibly read assay is set by controlling the number of accumulated visible labels to a number of labels which is less than can be seen with the unaided eye when the analyte present is at the desired limit of detection. As shown in the graph, there is a spectrum of the level of analyte in a particular sample, which will affect the reading of the test. When the number of labels is less than the limit of detection, the test line (120) will not be visible and the test result is interpreted as a negative test.
  • test line (150) will appear faintly as in graphic "A".
  • a control line (110) is preferably used in all tests to show that the test is working.
  • the number of false test readings can be reduced by combining 10% fluorescent labels with the visible labels.
  • Figure 7 shows a close up view of the "A-10% fluorescent labels" test result.
  • the fluorescent labels are preferably a fluorescing dye or fluorescing bead conjugates. When the analyte is at or near the limit of detection and the visible label is faint, the fluorescent labels will produce a visible signal when excited by a 490 nm diode laser. The visible line (130) of the fluorescing labels will confirm the test as a positive test (refer to graphic "A- 10% fluorescent labels").
  • Figure 8 shows a close-up view of another example of a positive test, where the fluorescent line (130) is shown.
  • the present invention provides for the reduction of interfering substances that might be present in the sample to be tested.
  • an interfering substance e.g. a human anti-mouse antibody (HAMA)
  • HAMA human anti-mouse antibody
  • the carrier may further include at least one capturing zone. Each capturing zone contains an immobilized capturing reagent specifically binding to a certain interfering substance, thereby immobilizing the interfering substance in the capturing zone.
  • the method allows a separation of the interfering substance or substances from the analyte or analytes of interest.
  • the capturing zone is located between the reagent zone and the detection zone.
  • the capturing zone may also be located between the application zone and the reagent zone.
  • the sample that has been collected is not lysed prior to collection and transfer to the sample analysis device. This decreases the number of steps needed to collect and prepare the sample for analysis.
  • the sample traveling with the transport liquid will encounter a lysis or lysing agent.
  • the lysis agent will have preferably been pre-loaded and dried onto the test strip and is eluted by the transport liquid.
  • the initially dried lysis agent is preferably localized between the sample application zone and a conjugate zone.
  • the lysis agent is preferably soluble in the sample transport liquid, and the lysis agent is solubilized and activated upon contact with the sample transport liquid.
  • the sample transport liquid then contains both lysis agent in solution or suspension and sample components in suspension. Any lysis- susceptible components in a sample, then being exposed in suspension to the lysis agent, are themselves lysed in situ.
  • the running buffer then carries the analyte, including any lysis-freed components, to a detection zone.
  • the location where the lysis agent is pre-loaded and dried can be varied as needed.
  • the dried lysis agent may be located in or just downstream of the sample application zone. Or, in order to minimize the distance along which the lysis product must travel before reaching the conjugate zone, the dried lysis agent may be located closer to the conjugate zone.
  • the present invention provides a lateral flow assay that uses the combined visual/fluorescence detection test to help differentiate viral and bacterial infections.
  • a combined point of care diagnostic device tests markers for both viral and bacterial infection, and can effectively assist in the rapid differentiation of viral and bacterial infections, for example at the outpatient office or during an urgent care visit. This ability can dramatically reduce health care costs by limiting misdiagnosis and the subsequent overuse of antibiotics. Such a practice may limit antibiotic allergies, adverse events, and antibiotic resistance.
  • the rapid result obtained from the test also permits a result while the patient is still being examined by the practitioner.
  • the marker for viral infection is MxA and the marker for bacterial infection is C-reactive protein (CRP).
  • CRP C-reactive protein
  • High MxA protein levels are strongly correlated with systemic viral infection and increased CRP is more associated with bacterial infections.
  • the present invention includes a rapid infectious screening test for identifying MxA and CRP in samples.
  • MxA is present in leukocytes (white blood cells). Therefore, the sample can be taken anywhere leukocytes are available, for example in a peripheral blood sample, nasopharyngeal aspirates, tears, spinal fluid, and middle ear aspirates.
  • markers for viral infection and/or bacterial infection may be used.
  • LCMV Lymphocytic Choriomeningitis Virus
  • Major transcription changes have been given preliminary confirmation by quantitative PCR and protein studies and are potentially valuable candidates as biomarkers for arenavirus disease.
  • markers for bacterial infection include, but are not limited to, procalcitonin, urinary trypsin inhibitor (uTi) , lipopolysaccharide, IL-I, IL-6, IL-8, IL-IO, ESR and an elevated WBC count (increased bands), Lactate, Troponin, vascular endothelial growth factor, platelet derived growth factor, Cortisol, proadrenomedullin, macrophage migratory inhibitory marker, activated protein C, CD 4,8,13,14, or 64, caspase, placenta derived growth factor, calcitonin gene-related peptide, high mobility group 1, copeptin, naturietic peptides, lipopolysaccharide binding protein, tumor necrosis factor alpha, circulating endothelial progenitor cells, complement 3a, and triggering receptor expresssed on myeloid cells (trem-1).
  • UTTi urinary trypsin inhibitor
  • the infections being distinguished are respiratory infections.
  • other types of infections which can be bacterial or viral, are differentiated using the system of the present invention.
  • Some examples include, but are not limited to, encephalitis, meningitis, gastroenteritis, febrile respiratory illness (including bronchitis, pharyngitis, pneumonia), sinusitis, otitis media, urinary tract infections, and conjunctivitis.
  • Red dyed 320 nm latex beads are coupled to 50 ug/ml AD 51 Adeno Hexon specific monoclonal antibody and 10 ug/ml of AD 51 Adeno Hexon specific monoclonal antibody is conjugated to the Dylight 405 fluorescing dye.
  • the conjugated beads are then blocked with 2% BSA containing 1 mg/ml Dylight 405 coupled BSA.
  • the visibly red latex beads conjugate also fluoresces under the UV light. This conjugate is used in a conjugate zone of the lateral flow test strip and serves as the visual (and fluorescing) tag in an Adeno DetectorTM lateral flow test.
  • Colloidal gold of 20 to 40 nm size conjugated to AD51 Adeno Hexon specific monoclonal antibody is mixed with colorless fluorescing latex beads conjugated to the same AD51 monoclonal antibody.
  • This mixture of visibly red colloidal gold conjugate and the colorless fluorescing latex beads is impregnated as the visual tag in the conjugate zone of an Adeno DetectorTM lateral flow strip.

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  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

Selon l'invention, la sensibilité de tests d'immunodosage à écoulement latéral et à lecture visuelle est améliorée par l'addition d'une petite quantité de colorant fluorescent ou de conjugués de particules de latex fluorescentes au matériau conjugué initial. Lorsque la ligne de test de spectre visible est visiblement présente, le résultat du test est observé et enregistré. Cependant, dans le cas où le résultat est indéterminé, une lumière d'un spectre approprié, telle qu'un spectre ultraviolet, visible ou infrarouge, est diffusée sur la ligne de test pour exciter et faire émettre une fluorescence aux particules de latex fluorescentes qui sont liées dans la ligne de test dans des tests vrais positifs, pour améliorer la couleur visible au niveau de la ligne de test.
PCT/US2009/046848 2008-06-10 2009-06-10 Test de détection d'analyte visuel/par fluorescence combiné WO2009152209A2 (fr)

Applications Claiming Priority (8)

Application Number Priority Date Filing Date Title
US6025808P 2008-06-10 2008-06-10
US61/060,258 2008-06-10
US8087908P 2008-07-15 2008-07-15
US61/080,879 2008-07-15
US9893508P 2008-09-22 2008-09-22
US61/098,935 2008-09-22
US17905909P 2009-05-18 2009-05-18
US61/179,059 2009-05-18

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WO2009152209A3 WO2009152209A3 (fr) 2010-03-11

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WO2013082943A1 (fr) * 2011-12-06 2013-06-13 Li Jiutong Analyse par fluorescence et dispositif associé
WO2013127144A1 (fr) * 2012-03-01 2013-09-06 上海鑫谱生物科技有限公司 Procédé et dispositif d'analyse par fluorescence
CN103293134A (zh) * 2012-03-01 2013-09-11 庞磊 一种荧光分析方法和装置
WO2014084260A1 (fr) * 2012-11-28 2014-06-05 古河電気工業株式会社 Immunochromatographie et détecteur et réactif pour utilisation dans celle-ci
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EP3591397A1 (fr) * 2013-03-07 2020-01-08 Rapid Pathogen Screening Inc. Procédé et dispositif de détection combinée d'infections virales et bactériennes
US10725033B2 (en) 2012-01-31 2020-07-28 Regents Of The University Of Minnesota Lateral flow assays with thermal contrast readers
US10808287B2 (en) 2015-10-23 2020-10-20 Rapid Pathogen Screening, Inc. Methods and devices for accurate diagnosis of infections
US10816492B2 (en) 2012-01-31 2020-10-27 Regents Of The University Of Minnesota Lateral flow assays with thermal contrast readers

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