Stabilized tetrazolium reagent compositions and methods for using the same

• US 6,586,199 B2
• Filed: 11/20/2001
• Issued: 07/01/2003
• Est. Priority Date: 11/20/2001
• Status: Expired due to Term

• Current Assignee: Cilag Gmbh International (Johnson & Johnson)
• Original Assignee: Lifescan Incorporated (Duv Holding Corporation)
• Inventors: Ouyang, Tianmei, Huang, Paing
• Primary Examiner(s): Leary, Louise N.
• Application Number:
  US09/988,812
• Publication Number:
  US 20030096332A1
• Time in Patent Office: 588 Days
• Field of Search: 435/26, 435/25, 435/14, 435/4, 435/975
• US Class Current: 435/26
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72

Abstract

Stabilized tetrazolium dye reagent compositions and methods for their use in the measurement of an analyte in a sample are provided. The subject reagent compositions include a tetrazolium dye component, e.g., a water soluble tetrazolium salt, and an effective amount of a nitrite stabilizing agent, e.g., a nitrite salt. In many embodiments, the subject reagent compositions include additional members of an analyte oxidizing signal producing system, such as: an analyte oxidizing enzyme, e.g., an analyte dehydrogenase or an analyte oxidase; an electron transfer agent; and an enzyme cofactor. Also provided are test strips that include the subject reagent compositions, as well as systems and kits incorporating the subject test strips. The subject reagent compositions, test strips, systems and kits find use in the detection of a wide variety of analytes in a sample, such as a physiological sample, e.g., blood or a fraction thereof, or ISF (interstitial fluid).

Claims

1. A reagent composition comprising:

a tetrazolium dye; and

an effective amount of a nitrite stabilizing agent.

2. The composition according to claim 1, wherein said tetrazolium dye is a water soluble tetrazolium salt.

3. The composition according to claim 1, wherein said nitrite stabilizing agent is a nitrite salt.

4. The composition according to claim 1, wherein said reagent composition comprises an analyte oxidizing signal producing system.

5. The composition according to claim 4, wherein said analyte oxidizing signal producing system comprises an analyte oxidase.

6. The composition according to claim 4, wherein said analyte oxidizing signal producing system comprises an analyte dehydrogenase.

7. The composition according to claim 4, wherein said analyte oxidizing signal producing system further comprises an electron transfer agent.

8. The composition according to claim 4, wherein said analyte oxidizing signal producing system further comprises an enzyme cofactor.

9. The composition according to claim 1, wherein said composition is a fluid composition.

10. The composition according to claim 1, wherein said composition is a dry composition.

17. The test strip according to claim 10, wherein said analyte oxidizing signal producing system is a glucose oxidizing signal producing system.

11. A reagent test strip comprising:

a substrate; and

an analyte oxidizing signal producing system present on said substrate, wherein said analyte oxidizing signal producing system includes a water soluble tetrazolium salt and a nitrite stabilizing agent.

12. The test strip according to claim 11, wherein said water soluble tetrazolium salt accepts a hydride to produce a water soluble formazan product.

13. The test strip according to claim 11, wherein said nitrite stabilizing agent comprises a nitrite salt.

14. The test strip according to claim 11, wherein said analyte oxidizing signal producing system comprises an analyte oxidase.

15. The test strip according to claim 14, wherein said analyte oxidizing signal producing system further comprises an electron transfer agent.

16. The test strip according to claim 14, wherein said analyte oxidizing signal producing system further comprises an enzyme cofactor.

18. An analyte detection or measurement system comprising:

(a) a reagent test strip comprising;

(i) a substrate; and

(ii) an analyte oxidizing signal producing system present on said substrate, wherein said signal producing system includes a water soluble tetrazolium salt capable of accepting a hydride to produce a water soluble formazan and a nitrite stabilizing agent; and

(b) an automated instrument.

19. A method for detecting the presence or determining the concentration of an analyte in a sample, said method comprising:

(a) applying said physiological sample to a reagent test strip comprising;

(i) a substrate; and

(ii) an analyte oxidizing signal producing system present on said substrate, wherein said signal producing system includes a water soluble tetrazolium salt capable of producing a water soluble formazan product and a nitrite stabilizing agent to produce a spot comprising said formazan product on said substrate;

(b) detecting said spot; and

(c) relating said detected spot to the presence or concentration of said analyte in said physiological sample.

20. The method according to claim 19, wherein said signal producing system further comprises an analyte oxidase.

21. The method according to claim 20, wherein said signal producing system further comprises at least one of an electron transfer agent.

22. The method according to claim 19, wherein said sample is whole blood or a derivative thereof.

23. The method according to claim 19, wherein said detecting and relating steps are carried out by an automated instrument.

24. A kit for use in determining the concentration of an analyte in a physiological sample, said kit comprising:

(a) a reagent test strip comprising;

(i) a substrate; and

(ii) an analyte oxidizing signal producing system present on said substrate, wherein said signal producing system includes a water soluble tetrazolium salt capable of producing a water soluble formazan product and a nitrite stabilizing agent; and

(b) at least one of;

(i) a means for obtaining said physiological sample and

(ii) an analyte standard.

25. The kit according to claim 24, wherein said means for obtaining said physiological sample is a lance.

26. The kit according to claim 24, wherein said analyte standard comprises a standardized concentration of a known reagent.

27. The kit according to claim 24, wherein said kit comprises a means for obtaining said physiological sample and an analyte standard.

28. A method for stabilizing a tetrazolium dye in a reagent composition, said method comprising:

including in said reagent composition an effective amount of a nitrite stabilizing agent.

29. The method according to claim 28, wherein said nitrite stabilizing agent is a nitrite salt.

30. The method according to claim 29, wherein said nitrite salt is sodium nitrite.

31. The method according to claim 28, wherein said reagent composition is a dry composition.

32. The method according to claim 28, wherein said reagent composition is a wet composition.