MICROFLUIDIC DEVICES AND METHODS FOR PATHOGEN DETECTION IN LIQUID SAMPLES

• US 20180104685A1
• Filed: 10/30/2017
• Published: 04/19/2018
• Est. Priority Date: 04/29/2015
• Status: Active Application

• Current Assignee: Administrators Of The Tulane Educational Fund (ADMINISTRATORS|DAVID H. COY)
• Original Assignee: Administrators Of The Tulane Educational Fund (ADMINISTRATORS|DAVID H. COY)
• Inventors: RYANS, Jason, SIVAKUMAR, Ashwin, DAVITT, Christopher
• Application Number:
  US15/797,102
• Publication Number:
  US 20180104685A1
• Time in Patent Office: Days
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/126 Paper
• CPC Class Codes: B01L 2300/165 Specific details about hydrophobic, oleophobic surfaces
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: G01N 2333/185 Flaviviruses or Group B arboviruses, e.g. yellow fever virus, japanese encephalitis, tick-borne encephalitis, dengue
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/569 for microorganisms, e.g. protozoa, bacteria, viruses
• CPC Class Codes: G01N 33/587 Nanoparticles

Abstract

One aspect of the present disclosure relates to a device for detecting a pathogen biomarker in a biological sample. The device can comprise a substrate. The substrate can comprise paper. The device can also comprise a hydrophobic material applied to the substrate to define at least one target region, a sample region, and at least one channel in fluid communication with the at least one target region and the sample region. The device can further include a predetermined amount of antibodies that specifically bind to at least one pathogen biomarker provided in the at least one target region. The predetermined amount of antibodies is conjugated with colloidal gold.

Claims

1. A microfluidic device comprising:

(a) a substrate;

(b) a hydrophobic material applied to the substrate to define at least one target region, a sample region for receiving a biological sample, and at least one channel in fluid communication with the at least one target region and the sample region; and

(c) a predetermined amount of antibodies that specifically bind to at least one pathogen biomarker provided in the at least one target region, wherein the predetermined amount of antibodies is conjugated with colloidal gold.

2. The device of claim 1, wherein the at least one pathogen biomarker comprises a pathogen antigen.

3. The device of claim 1, wherein the at least one pathogen biomarker comprises a mammalian antibody that specifically binds to a pathogen antigen.

4. The device of claim 1, wherein the antibodies comprise biotin conjugated antibodies and the colloidal gold comprises streptavidin conjugated colloidal gold.

5. The device of claim 4, wherein binding of the antibodies conjugated with colloidal gold to the at least one pathogen biomarker is indicated by a change in the visible color spectrum from red to purple.

6. The device of claim 1, wherein the substrate comprises paper.

7. The device of claim 1, wherein the hydrophobic material comprises wax.

8. The device of claim 1, further comprising a positive control.

9. The device of claim 8, wherein the positive control comprises colloidal gold particles conjugated with anti-human IgG or anti-human IgA.

10. The device of claim 1, further comprising a negative control.

11. The device of claim 1, further comprising a positive control and a negative control.

12. The device of claim 1, wherein the at least one pathogen biomarker comprises at least one viral pathogen biomarker.

13. The device of claim 12, wherein the at least one viral pathogen is selected from the group consisting of dengue-1 virus, dengue-2 virus, dengue-3 virus, dengue-4 virus, and combinations thereof.

14. The device of claim 1, wherein the viral pathogen is bovine viral diarrhea virus.

15. The device of claim 1, wherein the at least one pathogen related biomarker comprises at least one bacterial pathogen biomarker.

16. The device of claim 1, being configured as a multichannel device and further comprising:

two or more target regions, and two or more channels in fluid communication with the two or more target regions to the sample region.

17. A method of detecting the presence of at least one pathogen in a subject comprising:

(a) obtaining a biological sample from the subject;

(b) applying the biological sample to a microfluidic device comprising

(i) a substrate;

wherein the substrate comprises a sample area and a targeting area;

(ii) a hydrophobic material applied to the substrate to define at least one target region, a sample region for receiving a biological sample, and at least one channel in fluid communication with the at least one target region and the sample region; and

(iii) a predetermined amount of antibodies that specifically bind to at least one pathogen biomarker provided in the at least one target region, wherein the predetermined amount of antibodies is conjugated with colloidal gold; and

(c) determining the presence of the at least one pathogen biomarker in the biological sample, wherein the at least one pathogen biomarker, if present, specifically binds to the predetermined amount of antibodies conjugated with colloidal gold in the target region causing the formation of a colloidal gold conglomerate, and wherein the presence of a colloidal gold conglomerate is indicative of the presence of the at least one pathogen in the subject.

18. The method of claim 17, wherein the formation of the colloidal gold conglomerate is indicated by a change in the visible color spectrum from red to purple.

19. The method of claim 17, wherein the antibodies comprise biotin conjugated antibodies and the colloidal gold comprises streptavidin conjugated colloidal gold.

20. The method of claim 17, wherein the substrate comprises paper.

21. The method of claim 17, wherein the hydrophobic material comprises wax.

22. The method of claim 17, wherein the at least one pathogen biomarker comprises at least one viral pathogen biomarker.

23. The method of claim 22, wherein the at least one viral pathogen biomarker is selected from the group consisting of dengue-1 virus, dengue-2 virus, dengue-3 virus, dengue-4 virus, and combinations thereof.

24. The method of claim 22, wherein the viral pathogen comprises bovine viral diarrhea virus.

25. The method of claim 17, wherein the at least one pathogen biomarker comprises at least one bacterial pathogen biomarker.

26. The method of claim 17, wherein the biological sample is selected from the group comprising blood, urine, saliva, semen, perspiration, and mucus.

27. The method of claim 17, wherein the subject is domesticated livestock.

28. The method of claim 17, wherein the subject is a human.

29. The method of claim 17, wherein the device is configured as a multichannel device and further comprises two or more target regions, and two or more channels in fluid communication with the two or more target regions to the sample region.