Preparation of diagnostic test strips containing tetrazolium salt indicators

• US 5,360,595 A
• Filed: 08/19/1993
• Issued: 11/01/1994
• Est. Priority Date: 08/19/1993
• Status: Expired due to Term

• Current Assignee: Miles, Inc. (Bayer AG)
• Original Assignee: Miles, Inc. (Bayer AG)
• Inventors: Bell, Douglas E., Chu, Amy H.
• Primary Examiner(s): ALEXANDER, LYLE
• Application Number:
  US08/108,295
• Time in Patent Office: 439 Days
• Field of Search: 422/56, 422/58, 422/61, 436/164, 436/166
• US Class Current: 422/420
• CPC Class Codes: G01N 33/521 Single-layer analytical elements

Abstract

Disclosed is an improved procedure for the manufacture of a diagnostic test device comprising a matrix having uniformly dispersed a tetrazolium salt and a reagent system designed to convert the tetrazolium to its colored formazan upon contacting the matrix with a fluid containing an analyte whose presence and/or concentration is being sought. The procedure involves applying the tetrazolium salt to the matrix from its solution in an organic solvent and, after drying, applying the reagent system to the matrix from its aqueous solution to which has been added hexanesulfonate as wetting agent.

Claims

1. In the method of applying an indicator/reagent system to a matrix carrier, said indicator comprising a tetrazolium salt which, in the presence of the reagent system and a predetermined analyte will undergo a redox reaction resulting in its reduction to a colored formazan, the improvement which comprises:

a) contacting the matrix with a solution comprising the tetrazolium salt in an organic solvent and drying the matrix to thereby provide a matrix having the tetrazolium salt uniformly dispersed therein; and

b) contacting the matrix having the tetrazolium salt uniformly dispersed therein with an aqueous solution of the reagent system which solution contains an alkylsulfonate of 5-7 straight chain carbon atoms and drying the matrix to thereby deposit the reagent system onto the matrix without eluting a significant amount of the tetrazolium salt therefrom and provide a matrix having the tetrazolium salt and reagent system uniformly dispersed therein.

2. The method of claim 1 wherein the reagent system contains hexokinase, adenosine triphosphate, glucose-6-phosphate dehydrogenase, diaphorase and nicotinamideadenine dinucleotide.

3. The method of claim 1 wherein the tetrazolium salt is represented by the formula:

##STR7## wherein X.sup.⊖

is a counteranion, R¹ and R³ are substituted or unsubstituted phenyl groups and R³ is substituted or unsubstituted phenyl or 2-thiazolyl.

4. The method of claim 1 wherein the matrix material is an absorbant paper, a woven or nonwoven cloth, a glass fiber filter, or a polymeric membrane or film.

5. The method of claim 4 wherein the matrix material is a porous zwitterionic charged nylon fabric having pore sizes ranging from 0.45 to 0.8μ

.

6. The method of claim 1 wherein the alkyl sulfonate is present in the solution in a concentration of from 0.5% to 1.3% w/v.

7. The method of claim 6 wherein the alkanesulfonate is present in an amount of from 0.7% to 1.1% w/v.

8. A method of applying an indicator/reagent system to a matrix carrier of a porous zwitterionic charged nylon fabric, said indicator comprising a tetrazolium salt capable of undergoing a redox reaction resulting in its reduction to a colored formazan in the presence of the reagent system and a predetermined analyte which method comprises:

a) contacting the matrix with a solution comprising the tetrazolium salt in an organic solvent and drying the matrix to thereby provide a matrix having the tetrazolium salt dispersed therein; and

b) contacting the matrix having the tetrazolium salt dispersed therein with an aqueous solution of the reagent system comprising hexokinase, adenosine triphosphate, glucose-6-phosphate dehydrogenase, diaphorase and nicotinamideadenine dinucloetide which solution contains an alkylsulfonate of 5-7 straight chain carbon atoms and drying the matrix to thereby deposit the reagent system onto the matrix while reducing the amount of tetrazolium salt which is eluted from the matrix from the aqueous solution.

9. The method of claim 8 wherein the alkanesulfonate is hexane sulfonate and is present in the aqueous solution in a concentration of from 0.5% to 1.3% w/v.

10. The method of claim 9 wherein the hexane sulfonate is present in the aqueous solution in a concentration of from 0.7% to 1.1% w/v.

11. A diagnostic test device prepared by:

a) contacting a matrix carrier with a solution comprising a tetrazolium salt which, in the presence of a reagent system and a predetermined analyte will undergo a redox reaction resulting in its reduction to a colored formazan in an organic solvent and drying the matrix to thereby provide a matrix having the tetrazolium salt uniformly dispersed therein; and

b) contacting the matrix having the tetrazolium salt uniformly dispersed therein with an aqueous solution of the reagent system which solution contains an alkylsulfonate of 5-7 straight chain carbon atoms and drying the matrix to thereby deposit the reagent system onto the matrix to provide a matrix having the tetrazolium salt and reagent system uniformly dispersed therein.

12. The device of claim 11 wherein the reagent system contains hexokinase, adenosine triphosphate, glucose-6-phosphate dehydrogenase, diaphorase and nicotinamideadenine dinucleotide.

13. The device of claim 11 wherein the tetrazolium salt is represented by the formula:

##STR8## wherein X.sup.⊖

is a counteranion, R¹ and R³ are substituted or unsubstituted phenyl groups and R³ is substituted or unsubstituted phenyl or 2-thiazolyl.

14. The device of claim 11 wherein the matrix material is an absorbent paper, a woven or nonwoven cloth, a glass fiber filter, or a polymeric membrane or film.

15. The device of claim 11 wherein the matrix material is a porous zwitterionic charged nylon fabric having pore sizes ranging from 0.45 to 0.8μ

.

16. The device of claim 11 wherein the alkyl sulfonate is present in the solution in a concentration of from 0.5% to 1.3% w/v.

17. The device of claim 11 wherein the alkanesulfonate is present in an amount of from 0.7% to 1.1% w/v.