Method and device for combined detection of viral and bacterial infections

• US 10,379,121 B2
• Filed: 05/25/2016
• Issued: 08/13/2019
• Est. Priority Date: 05/20/2008
• Status: Active Grant

• Current Assignee: Rapid Pathogen Screening, Inc
• Original Assignee: Rapid Pathogen Screening, Inc
• Inventors: Sambursky, Robert P., VanDine, Robert W., Babu, Uma Mahesh, Condon, Peter
• Primary Examiner(s): Crawford, Erik B
• Application Number:
  US15/164,137
• Publication Number:
  US 20160266119A1
• Time in Patent Office: 1,175 Days
• Field of Search: None
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00648 by the use of solid beads
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/0074 Biological products
• CPC Class Codes: B01J 2219/00743 Cells
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: G01N 2333/4703 Regulators; Modulating activity
• CPC Class Codes: G01N 2333/4737 C-reactive protein
• CPC Class Codes: G01N 2333/914 Hydrolases (3)
• CPC Class Codes: G01N 2469/00 Immunoassays for the detection of microorganisms
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/569 for microorganisms, e.g. protozoa, bacteria, viruses
• CPC Class Codes: G01N 33/56911 Bacteria
• CPC Class Codes: G01N 33/56983 Viruses

Abstract

A lateral flow assay is capable of detecting and differentiating viral and bacterial infections. A combined point of care diagnostic device tests markers for viral infection and markers for bacterial infection, to effectively assist in the rapid differentiation of viral and bacterial infections. In some preferred embodiments, bimodal methods and devices determine if an infection is bacterial and/or viral. A dual use two strip sample analysis device includes a first lateral flow chromatographic test strip to detect MxA and a low level of C-reactive protein and a second lateral flow chromatographic test strip to detect high levels of C-reactive protein. In some preferred embodiments, the sample is a fingerstick blood sample.

Claims

1. A method for analyzing a sample for a presence of MxA and C-reactive protein, comprising the steps of:

a) collecting a fresh, fingerstick whole blood sample;

b) transferring the sample to a sample analysis device comprising;

i) a first lateral flow chromatographic test strip comprising;

a first reagent zone for detecting a low level intensity of C-reactive protein comprising at least one first reagent specific to C-reactive protein such that, when the sample contacts the first reagent, a first labeled complex forms if the low level intensity of C-reactive protein is present is greater than a threshold concentration in the sample;

a second reagent zone comprising at least one second reagent specific to detect an intensity MxA such that, when the sample contacts the second reagent, a second labeled complex forms if MxA is present in the sample; and

a first detection zone comprising a first binding partner which binds to the first labeled complex indicative of detecting a bacterial marker being in the sample, wherein the threshold concentration to obtain a positive visual result for the low level intensity of C-reactive protein in the first detection zone of the first lateral flow chromatographic test strip is equal to or greater than a serum equivalent of approximately 6-15 mg/L of C-reactive protein; and

a second binding partner which binds to the second labeled complex indicative of detecting a viral marker being present in the sample;

an

ii) a second lateral flow chromatographic test strip parallel in a lateral flow direction to the first lateral flow chromatographic test strip, comprising;

at least one third reagent zone for detecting a high level intensity of C-reactive protein comprising;

at least one third reagent specific to C-reactive protein, wherein the third reagent only detects a level of intensity C-reactive protein that is higher in intensity than the low level of intensity of C-reactive protein detected by the first reagent, such that, when the sample contacts the third reagent, a third labeled complex forms if the high level intensity of C-reactive protein is present in the sample; and

a second detection zone comprising a third binding partner which binds to the third labeled complex wherein the threshold concentration to obtain a positive result for the high level of intensity of C-reactive protein in the second reagent zone of the second lateral flow chromatographic test strip is equal to or greater than a serum equivalent of approximately 60-100 mg/L;

wherein the second lateral flow chromatographic test strip does not comprise reagent and detection zones for detecting MxA; and

c) analyzing the sample for a presence of the low level intensity of C-reactive protein, MxA, and the high level intensity of C-reactive protein;

wherein the fresh fingerstick whole blood sample is transferred to the sample analysis device and analyzed, such that a result is obtained in 20 minutes or less.

2. The method of claim 1, wherein the threshold concentration to obtain a positive result for the low level intensity of C-reactive protein in the first detection zone of the first lateral flow chromatographic test strip is equal to or greater than a serum equivalent of approximately 10 mg/L of C-reactive protein.

3. The method of claim 1, wherein a threshold concentration to obtain a positive result for the high level intensity of C-reactive protein in the second detection zone of the second lateral flow chromatographic test strip is equal to or greater than a serum equivalent of approximately 80 mg/L.

4. The method of claim 1, wherein a threshold concentration to obtain a positive result for MxA in the first detection zone of the first lateral flow chromatographic test strip is equal to or greater than approximately 40 ng/ml.

5. The method of claim 1, wherein a threshold concentration to obtain a positive result for MxA in the first detection zone of the first lateral flow chromatographic test strip is equal to or greater than approximately 15-250 ng/ml.

6. The method of claim 1, wherein a first threshold concentration to obtain a positive result for the low level intensity of C-reactive protein in the first detection zone of the first lateral flow chromatographic test strip is equal to or greater than a serum equivalent of approximately 10 mg/L of C-reactive protein, a second threshold concentration to obtain a positive result for the high level intensity of C-reactive protein in the second detection zone of the second lateral flow chromatographic test strip is equal to or greater than a serum equivalent of approximately 80 mg/L, and a third threshold concentration to obtain a positive result for MxA in the first detection zone of the first lateral flow chromatographic test strip is equal to or greater than approximately 40 ng/ml.

7. The method of claim 1, wherein step c) comprises the substeps of:

i) eluting the sample on the sample analysis device; and

ii) visually determining a result from the first and/or second detection zone.

8. The method of claim 1, wherein the presence of MxA is indicated by a first test line located in the first detection zone of the first lateral flow chromatographic test strip and the presence of the low level intensity of C-reactive protein is indicated by a second test line located in the first detection zone of the first lateral flow chromatographic test strip.

9. The method of claim 8, wherein the first test line displays a first color when positive and the second test line displays a second color different from the first color when positive.

10. The method of claim 9, wherein both the first test line and the second test line are located in the same space on the sample analysis device such that a third color is formed when both the first test line and the second test line are positive.

11. The method of claim 8, wherein the first test line is spatially separate from the second test line on the first lateral flow chromatographic test strip.

12. The method of claim 1, wherein the sample contains leukocytes.

13. The method of claim 1, wherein the results are obtained in 10 minutes or less.