Method and device for combined detection of viral and bacterial infections

• US 8,962,260 B2
• Filed: 03/08/2013
• Issued: 02/24/2015
• 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): Brown, Melanie Y
• Assistant Examiner(s): Crawford, Erik B
• Application Number:
  US13/790,160
• Publication Number:
  US 20130196311A1
• Time in Patent Office: 718 Days
• Field of Search: 435/5, 435/7.1, 435/7.2, 435 79- 795, 436/810, 436/514, 436/518, 436524-532, 436536-541
• US Class Current: 435/7.1
• 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 21/8483 Investigating reagent band test-element handling not specific to a test method G01N33/4875; analytical elements specific to chemical analysis of biological material G01N33/52; autometer with reagent band G01N35/04
• CPC Class Codes: G01N 30/90 Plate chromatography, e.g. thin layer or paper chromatography
• CPC Class Codes: G01N 33/523 the element being adapted for a specific analyte
• CPC Class Codes: G01N 33/526 the element being adapted for a specific analyte
• CPC Class Codes: G01N 33/54346 Nanoparticles
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/54389 with bidirectional or multidirectional lateral flow, e.g. wherein the sample flows from a single, common sample application point into multiple strips, lanes or zones
• 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
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

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 sample;

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

i) a sample compressor comprising;

A) a first reagent zone for detecting a low level of C-reactive protein comprising at least one first reagent specific to of C-reactive protein such that, when the sample contacts the first reagent, a first labeled complex forms if the low level of C-reactive protein is present in the sample and at least one second reagent specific to MxA such that, when the sample contacts the second reagent, a second labeled complex forms if MxA is present in the sample; and

B) a second reagent zone for detecting a high level of C-reactive protein comprising at least one third reagent specific to C-reactive protein, wherein the third reagent only detects a level of C-reactive protein that is higher than the level 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 of C-reactive protein is present in the sample;

ii) a first lateral flow chromatographic test strip comprising;

A) a first detection zone comprising a first binding partner which binds to the first labeled complex; and

a second binding partner which binds to the second labeled complex; and

B) a first diverting zone located upstream of the first detection zone on the lateral flow chromatographic test strip, wherein the first diverting zone interrupts lateral flow on the first lateral flow chromatographic test strip; and

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

A) a second detection zone comprising a third binding partner which binds to the third labeled complex; and

B) a second diverting zone located upstream of the second detection zone on the second lateral flow chromatographic test strip, wherein the diverting zone interrupts lateral flow on the second lateral flow chromatographic test strip;

iv) a first sample application zone where sample is placed on the sample analysis device, wherein the first sample application zone is located in a location selected from the group consisting of;

i) on the first lateral flow chromatographic test strip upstream of the detection zone and ii) on the first reagent zone of the sample compressor; and

v) a second sample application zone where sample is placed on the sample analysis device, wherein the second sample application zone is located in a location selected from the group consisting of;

i) on the second lateral flow chromatographic test strip upstream of the detection zone and ii) on the second reagent zone of the sample compressor;

wherein the sample compressor is in a different plane than the first lateral flow chromatographic test strip and the second lateral flow chromatographic test strip;

wherein the first reagent zone of the sample compressor creates a bridge over the first diverting zone and the second reagent zone of the sample compressor creates a bridge over the second diverting zone, diverting flow onto the sample compressor and returning flow to the first chromatographic test strip and the second chromatographic test strip at downstream ends of the first diverting zone and the second diverting zone; and

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

2. The method of claim 1 wherein the sample analysis device further comprises a first control binding partner located on each of the first reagent zone and the second reagent zone of the sample compressor and a second control binding partner immobilized in a control zone of each of the first lateral flow chromatographic test strip and the second lateral flow chromatographic test strip, wherein the first control binding partner is a binding partner for the second control binding partner.

3. The method of claim 1, wherein a threshold concentration to obtain a positive result for the low level of C-reactive protein in the 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.

4. The method of claim 1, wherein the threshold concentration to obtain a positive result for the low level of C-reactive protein in the 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.

5. The method of claim 1, wherein a threshold concentration to obtain a positive result for the high level of C-reactive protein in the detection zone of the second lateral flow chromatographic test strip is equal to or greater than a serum equivalent of approximately 60-100 mg/L.

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

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

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

9. The method of claim 1, wherein a first threshold concentration to obtain a positive result for the low level of C-reactive protein in the 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, a second threshold concentration to obtain a positive result for the high level of C-reactive protein in the detection zone of the second lateral flow chromatographic test strip is equal to or greater than a serum equivalent of approximately 60-100 mg/L, and a third threshold concentration to obtain a positive result for MxA in the detection zone of the first lateral flow chromatographic test strip is equal to or greater than approximately 15-250 ng/ml.

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

11. 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 zones.

12. 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 of CRP is indicated by a second test line located in the first detection zone of the first lateral flow chromatographic test strip.

13. The method of claim 12, 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.

14. The method of claim 13, 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.

15. The method of claim 12, wherein the first test line is spatially separate from the second test line on the first lateral flow chromatographic test strip.

16. The method of claim 1, wherein the sample is a blood sample.

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

41. The method of claim 1, further comprising a step of determining whether an infection is bacterial and/or viral based on the presence of MxA and/or C-reactive protein in the sample.

18. A lateral flow device for detecting an analyte in a sample comprising:

a) a sample compressor comprising;

i) a first reagent zone 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 of C-reactive protein is present in the sample and at least one second reagent specific to MxA such that, when the sample contacts the second reagent, a second labeled complex forms if MxA is present in the sample; and

ii) a second reagent zone for detecting a high level of C-reactive protein comprising at least one third reagent specific to C-reactive protein, wherein the third reagent only detects a level of C-reactive protein that is higher than the level of C-reactive protein detected by the second reagent, such that, when the sample contacts the third reagent, a third labeled complex forms if the high level of C-reactive protein is present in the sample;

b) a first lateral flow chromatographic test strip comprising;

i) a first detection zone comprising a first binding partner which binds to the first labeled complex; and

a second binding partner which binds to the second labeled complex; and

ii) a first diverting zone located upstream of the first detection zone on the lateral flow chromatographic test strip, wherein the first diverting zone interrupts lateral flow on the first lateral flow chromatographic test strip; and

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

i) a second detection zone comprising a third binding partner which binds to the third labeled complex; and

ii) a second diverting zone located upstream of the first detection zone on the lateral flow chromatographic test strip, wherein the second diverting zone interrupts lateral flow on the second lateral flow chromatographic test strip; and

d) a first sample application zone where sample is placed on the sample analysis device, wherein the first sample application zone is located in a location selected from the group consisting of;

i) on the first lateral flow chromatographic test strip upstream of the detection zone and ii) on the first reagent zone of the sample compressor;

e) a second sample application zone where sample is placed on the sample analysis device, wherein the second sample application zone is located in a location selected from the group consisting of;

i) on the second lateral flow chromatographic test strip upstream of the detection zone and ii) on the second reagent zone of the sample compressor;

wherein the sample compressor is in a different plane than the first lateral flow chromatographic test strip and the second lateral flow chromatographic test strip;

wherein the first reagent zone of the sample compressor creates a bridge over the first diverting zone and the second reagent zone of the sample compressor creates a bridge over the second diverting zone, diverting flow onto the sample compressor and returning flow to the first chromatographic test strip and the second chromatographic test strip at downstream ends of the first diverting zone and the second diverting zone.

19. The device of claim 18 wherein the device further comprises a first control binding partner located on each of the first reagent zones and the second reagent zones of the sample compressor and a second control binding partner immobilized in a control zone of each of the first lateral flow chromatographic test strip and the second lateral flow chromatographic test strip, wherein the first control binding partner is a binding partner for the second control binding partner.

20. The device of claim 18, wherein a threshold concentration to obtain a positive result for the low level 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.

21. The device of claim 18, wherein the threshold concentration to obtain a positive result for the low level 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.

22. The device of claim 18, wherein a threshold concentration to obtain a positive result for the high level 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 60-100 mg/L.

23. The device of claim 18, wherein a threshold concentration to obtain a positive result for the high level 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.

24. The device of claim 18, 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.

25. The device of claim 18, 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.

26. The device of claim 18, wherein a first threshold concentration to obtain a positive result for the low level 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, a second threshold concentration to obtain a positive result for the high level 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 60-100 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 15-250 ng/ml.

27. The device of claim 18, wherein a first threshold concentration to obtain a positive result for the low level 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 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.

28. The device of claim 18, wherein the first detection zone of the first lateral flow chromatographic test strip comprises a first test line to detect a positive result for MxA in the sample and a second test line to detect a positive result for the low level of CRP in the sample.

29. The device of claim 28, 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.

30. The device of claim 29, wherein both the first test line and the second test line are located in the same space on the first lateral flow chromatographic test strip such that a third color is formed when both the first test line and the second test line are positive.

31. The device of claim 28, wherein the first test line is spatially separate from the second test line on the first lateral flow chromatographic test strip.

32. The device of claim 18, wherein the first detection zone and the second detection zone each comprise a control line that is visible to the naked eye when the device is working.

33. The device of claim 18, wherein the first lateral flow chromatographic test strip further comprises a first sample application zone upstream of the first reagent zone, the second reagent zone, and the first detection zone, and the first detection zone is downstream of the first reagent zone and the second reagent zone.

34. The device of claim 18, wherein the second lateral flow chromatographic test strip further comprises a second sample application zone upstream of the third reagent zone, and the second detection zone, and the second detection zone is downstream of the third reagent zone.

35. The device of claim 18, wherein the first lateral flow chromatographic test strip further comprises a lysis zone comprising at least one lysis agent, wherein the lysis agent contacts the sample on the first lateral flow chromatographic test strip.

36. The device of claim 18, wherein the second lateral flow chromatographic test strip further comprises a lysis zone comprising at least one lysis agent, wherein the lysis agent contacts the sample on the second lateral flow chromatographic test strip.

37. The device of claim 18, wherein the first lateral flow chromatographic test strip further comprises a first sample application zone downstream of the first reagent zone and the second reagent zone, and upstream the first detection zone.

38. The device of claim 18, wherein the second lateral flow chromatographic test strip further comprises a second sample application zone downstream of the third reagent zone and upstream of the second detection zone.

39. The device of claim 18, wherein the sample is a blood sample.

40. The device of claim 18, wherein the sample contains leukocytes.