Immunoassay for venom detection including noninvasive sample collection

• US 8,426,154 B2
• Filed: 04/08/2010
• Issued: 04/23/2013
• 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, The United States of America As Represented By The Secretary of The Air Force Directorate of Intellectual Property Law, University of Michigan, Curators of the University of Missouri (University of Missouri)
• Inventors: Stoecker, William V., Gomez, Hernan F., Green, Jonathan A., McGlasson, David L.
• Primary Examiner(s): Haq, Shafiqul
• Assistant Examiner(s): YAKOVLEVA, GALINA M
• Application Number:
  US12/756,875
• Publication Number:
  US 20100196938A1
• Time in Patent Office: 1,111 Days
• Field of Search: None
• US Class Current: 435/7.92
• 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 Loxosceles spider bite in a patient, said method comprising:

(a) collecting a sample comprising venom from the area of the bite using a swab;

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

(c) detecting a complex formed by binding of said antibody to said venom with a tracer.

2. The method of claim 1 wherein said swab comprises an absorbent or adsorbent material selected from the group consisting of cotton, synthetic fibers, 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 Loxosceles spider is a Loxosceles Reclusa.

11. 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.

12. The method of claim 11 wherein said ELISA is capable of identifying under 24 picograms of venom in a sample.

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

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