IMMUNOASSAY FOR VENOM DETECTION INCLUDING NONINVASIVE SAMPLE COLLECTION

• US 20070141626A1
• Filed: 10/17/2006
• Published: 06/21/2007
• Est. Priority Date: 10/17/2005
• Status: Active Grant

• Current Assignee: Spidertech Inc., United States Of America As Represented By The Secretary Of The Air Force
• Original Assignee: Spidertech Inc., United States Of America As Represented By The Secretary Of The Air Force, Curators of the University of Missouri (University of Missouri)
• Inventors: Green, Jonathan, Gomez, Hernan, Stoecker, William, McGlasson, David
• Granted Patent: US 7,927,828 B2
• Time in Patent Office: Days
• US Class Current: 435/7.100
• CPC Class Codes: G01N 2800/42 Poisoning, e.g. from bites or stings
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

Abstract

Methods and immunoassays for diagnosing a bite or sting of a venomous organism in a patient having symptoms consistent with such a bite or sting are provided. A sample of venom is collected from the area of the suspected bite or sting using a swab and then contacted with an antibody that specifically binds to an antigenic site on venom present in the sample. Binding is then detected. The invention is illustrated by examples showing diagnosis of brown recluse spider bite, distinguishing it from other diagnoses with which it is often confused. This extremely sensitive test can detect venom antigens down to about 20 picograms even after the sample has been shipped and stored for periods of up to three weeks during the summer.

Claims

1. A method of diagnosing a bite or sting of a venomous organism in a patient, said method comprising:

(a) collecting a sample comprising venom from said venomous organism from the area of the bite or sting using a swab;

(b) contacting the sample with an antibody which specifically binds to an antigenic site on venom present in the sample; and

(c) detecting a complex formed by binding of said antibody and said antigenic site.

2. The method of claim 1 wherein said swab comprises an absorbent or adsorbent material selected from the group consisting of cotton, Dacron, rayon, polyurethane foam, brushes, paper and sponge.

3. The method of claim 1 wherein said detecting is performed by a method selected from the group consisting of:

sandwich immunoassays and electrochemical immunoassays.

4. The method of claim 1 wherein said detecting is performed using an immunoassay device outside a laboratory.

5. The method of claim 1 wherein said swab also comprises said antibody.

6. The method of claim 1 wherein said swab is disposable.

7. The method of claim 1 wherein said sample is collected by gently wiping or soaking the skin with said swab for about one to about 360 seconds.

8. The method of claim 1 also comprising collecting a control sample from a separate site on the patient'"'"'s body that has not been exposed to said venom.

9. The method of claim 1 wherein said sample comprises blister fluid.

10. The method of claim 1 wherein said venomous organism is selected from the group consisting of venomous spiders, venomous scorpions, venomous snakes, venomous wasps, venomous bees, and venomous jellyfish.

11. The method of claim 1 wherein said venomous organism is a spider selected from the genuses Latrodectus, Loxosceles, Atrax, Hadronyche, Tegenaria and Phoneutria or the family Theraphosidae.

12. The method of claim 11 wherein said venomous organism is a Loxosceles Reclusa.

13. The method of claim 1 wherein said venomous organism is a jellyfish.

14. The method of claim 1 wherein said venomous organism is a scorpion.

15. The method of claim 1 wherein said venomous organism is a snake.

16. The method of claim 1 wherein said detecting is performed using an ELISA modified to reduce blocking antibodies using nonfat milk solids and alkaline phosphatase markers.

17. The method of claim 16 wherein said ELISA is capable of identifying under 24 picograms of venom in a sample.

18. The method of claim 1 wherein said sample is exposed to ambient summertime temperatures for up to about three weeks prior to said detecting.

19. The method of claim 1 wherein said detecting distinguishes said organism from other species of venomous organism.

20. An immunoassay kit comprising:

(a) at least one antivenom antibody capable of binding to an antigen present in a venom;

(b) a swab for collecting a sample comprising venom from the area of a venomous bite or sting on a patient'"'"'s body;

(c) a tracer for detecting binding between said antibody and said antigen.

21. The immunoassay kit of claim 20 wherein said antibody is immobilized on a solid substrate.

22. The immunoassay kit of claim 21 wherein said solid substrate is said swab.

23. The immunoassay kit of claim 20 wherein said swab comprises an absorbent or adsorbent material selected the group consisting of cloth, gauze, sponge, polymeric foam, cotton, rayon, Dacron, or nylon fibers, polyurethane foam, paper, and absorbent or adsorbent brushes.

24. The immunoassay kit of claim 20 wherein said swab is attached to a handle.

25. The immunoassay kit of claim 20 wherein said swab comprises natural or synthetic fibers.

26. The immunoassay kit of claim 20 wherein said swab comprises bristles.

27. The immunoassay kit of claim 20 wherein at least one antivenom antibody is a polyclonal antibody.

28. The immunoassay kit of claim 27 wherein said polyclonal antibody has been raised in a rabbit.

29. The immunoassay kit of claim 20 wherein said tracer comprises a monoclonal antibody capable of binding to said antivenom antibody.

30. The immunoassay kit of claim 20 wherein said immunoassay is a portable calorimetric assay.

31. The immunoassay kit of claim 20 wherein said antivenom antibody is immobilized on an absorbent support.

32. The immunoassay kit of claim 20 also comprising a label attached to said tracer that indicates binding of said antivenom antibody to said venom protein wherein said label is selected from the group consisting of including enzymes, radioactive isotopes, and color particles.

33. The immunoassay kit of claim 32 also comprising detector means for detecting the presence of said label or changes in said label that indicate the tracer has bound to the antivenom antibody.

34. The immunoassay kit of claim 20 also comprising means for lysing cells present in said sample.

35. The immunoassay kit of claim 20 wherein said antivenom antibody is an antibody against a protein component of Loxosceles reclusa venom.

36. The immunoassay kit of claim 20 wherein said antivenom antibody is an antibody against sphingomyelinase.

37. The immunoassay kit of claim 20 also comprising instructions for use thereof.