Dip-stick assay for C-reactive protein

• US 6,406,862 B1
• Filed: 10/06/1999
• Issued: 06/18/2002
• Est. Priority Date: 10/06/1998
• Status: Expired due to Fees

• Current Assignee: United States Army (U.S. Department Of Defense)
• Original Assignee: The United States Of America As Represented By The Secretary Of The Army
• Inventors: Krakauer, Teresa
• Primary Examiner(s): Stucker, Jeffrey
• Assistant Examiner(s): JENKS, ULRIKE WINKLER
• Application Number:
  US09/413,361
• Time in Patent Office: 986 Days
• Field of Search: 435/4, 435/7.1, 435/7.5, 435/7.6, 435/7.92, 435/7.93, 435/7.94, 435/805, 436/513, 436/518, 530/380, 530/388.1, 530/389.1
• US Class Current: 435/7.1
• CPC Class Codes: G01N 2333/4737 C-reactive protein
• CPC Class Codes: G01N 33/6893 related to diseases not provided for elsewhere
• CPC Class Codes: Y10S 435/805 Test papers

Abstract

A C-reactive protein concentration level test and kit for on-site determination of C-reactive protein levels in biological samples is disclosed.

Claims

1. A method for detecting tissue injury, inflammation or infection in a subject comprising measuring levels of C-reactive protein in a biological sample of said subject, comprising

(i) binding an antibody against CRP to a solid support chosen from the group consisting of dip-stick and membrane;

(ii) incubating said solid support in the presence of the sample to be analyzed under conditions where antibody-antigen complexes form;

(iii) incubating said support with an anti-CRP antibody conjugated to a detectable moeity which produces a signal;

(vi) visually detecting said signal wherein said signal is proportional to the amount of CRP in said sample; and

(v) comparing the signal in said sample to a standard wherein a level equal to or below 10 mg/l is considered negative, and a level above 10 mg/l is considered indicative of tissue injury, infection or inflammation in said subject.

2. The method of claim 1 wherein said biological sample is serum.

3. The method of claim 1 wherein the standard is assayed simultaneously along with the biological sample.

4. The method of claim 1 wherein said standards range from about 10 mg of CRP per liter of solution to about 100 mg of CRP per liter of solution.

5. The method of claim 1 wherein said detection is by use of a chromogen.

6. The method according to claim 5 wherein said chromogen, is 3,3′

5,5′

-tetramethylbenzidine or its derivative.

7. The assay according to claim 1 wherein said assay is performed at room temperature.

8. The assay according to claim 1 wherein said biological sample is 5 microliters.

9. The method according to claim 1 wherein said method is performed at room temperature.

10. A test kit for visual detection of C-reactive protein wherein detection of a level equal to or below 10 mg/l is considered negative, and a level above 10 mg/l is considered indicative of tissue injury, infection or inflammation in said subject, said kit comprising:

one or more solutions containing a known concentration of CRP to serve as a standard;

a solution of a anti-CRP antibody bound to an enzyme;

a chromogen which changes color or shade by the action of the enzyme;

a solid support chosen from the group consisting of dip-stick and membrane carrying on the surface thereof an antibody to C-reactive protein.

11. A test kit of claim 10 wherein said kit further comprises test tubes for said solutions.