Test strip for detection of analyte and methods of use

• US 20030232321A1
• Filed: 06/15/2002
• Published: 12/18/2003
• Est. Priority Date: 06/15/2002
• Status: Active Grant

• Current Assignee: Leadway (Hk) Limited (Akatsuki Corp.)
• Original Assignee: Oakville Trading Hong Kong Limited
• Inventors: Wang, Chia-Lin, Lin, Jinn-Nan
• Granted Patent: US 6,759,190 B2
• Time in Patent Office: Days
• US Class Current: 435/4
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: Y10S 435/968 High energy substrates, e.g. fluorescent, chemiluminescent, radioactive
• CPC Class Codes: Y10S 435/97 Test strip or test slide

Abstract

One aspect of the present invention is a reagent test strip that includes: a) a top support layer including a sample aperture; b) a membrane that includes a reagent system for indicating the concentration of a target; c) a spreading layer; d) a bottom support layer including a measuring port in substantial alignment or approximate alignment with the sample aperture. Preferably, the membrane is affixed to the top support layer. Preferably, the membrane is positioned between the top support layer and the bottom support layer. Preferably, the spreading layer is positioned above the top support layer and in substantial or approximate alignment with the sample aperture. Preferably, a fluid applied to the spreading layer, passes through the sample aperture and contacts the membrane.

Claims

1. A reagent test strip comprising:

a. a top support layer comprising a sample aperture;

b. a membrane comprising a reagent system for indicating the concentration of a target;

c. a spreading layer;

d. a bottom support layer comprising a measuring port in substantial alignment or approximate alignment with said sample aperture;

wherein said membrane is affixed to said top support layer;

further wherein said membrane is positioned between said top support layer and said bottom support layer;

further wherein said spreading layer is positioned above said top support layer and in substantial or approximate alignment with said sample aperture; and

further wherein a fluid applied to said spreading layer, passes through said sample aperture and contacts said membrane.

2. A reagent test strip according to claim 1 wherein said membrane is hydrophilic.

3. A reagent test strip according to claim 1 wherein said membrane is able to substantially exclude red blood cells.

4. A reagent test strip according to claim 1 wherein said membrane is a symmetric membrane or an asymmetric membrane.

5. A reagent test strip according to claim 1 wherein said membrane comprises polysulfone, polyethersulfone, nylon, or nitrocellulose.

6. A reagent test strip according to claim 1 wherein said reagent system comprises an enzyme.

7. A reagent test strip according to claim 6 wherein said enzyme is glucose oxidase.

8. A reagent test strip according to claim 6 wherein said enzyme is horseradish peroxidase.

9. A reagent test strip according to claim 1 wherein said reagent system comprises glucose oxidase and horseradish peroxidase.

10. A reagent test strip according to claim 1 wherein said reagent system comprises a substrate capable of forming a chromophore

11. A reagent test strip according to claim 10 wherein said substrate is a AAP and MAOS.

12. A reagent test strip according to claim 10 wherein said chromophore is detected by measuring reflectance.

13. A reagent test strip according to claim 12 wherein said reflectance is indicative of the concentration of said target.

14. A reagent test strip according to claim 12 wherein said reflectance is detected directly on said membrane.

15. A reagent test strip according to claim 10 wherein said chromophore absorbs light from about 500 nm to about 800 nm.

16. A reagent test strip according to claim 10 wherein said chromophore absorbs light from about 600 nm to about 700 nm.

17. A reagent test strip according to claim 1 wherein said target is a biological moiety or chemical moiety.

18. A reagent test strip according to claim 1 wherein said target is a protein or nucleic acid.

19. A reagent test strip according to claim 1 wherein said target is glucose.

20. A reagent test strip according to claim 1 wherein said spreading layer is hydrophillic.

21. A reagent test strip according to claim 1 wherein said fluid comprises whole blood.

22. A reagent test strip according to claim 1, wherein said test strip reduces wet through of a said fluid through said membrane.

24. A method of determining the presence or concentration of a target in a sample comprising the steps of:

a. applying a fluid suspected of comprising a target to the reagent test strip of claim 1;

b. detecting the reflectance of said membrane through said measuring port; and

c. determining the presence or concentration of said target.

25. A method of determining the presence or concentration of a target in a sample according to claim 24 wherein the step of determining the presence or concentration comprises measuring the absorbance of a chromophore.

26. A method of determining the concentration of a target in a sample comprising the steps of:

a. detecting a dry strip reading from the reagent test strip of claim 1;

b. running a preprogrammed threshold loop instruction;

c. detecting a first reflectance measurement;

d. running a preprogrammed kinetic loop instruction;

e. determining the presence or concentration of the target;

wherein said threshold loop instruction comprises the steps of;

detecting a threshold comparison measurement;

comparing said threshold comparison measurement to a predetermined threshold value;

repeating the loop until said threshold comparison measurement is less than or equal to said predetermined threshold value;

further wherein a fluid suspected of including a target is applied during the step of running said threshold loop instruction;

further wherein said kinetic loop instruction comprises the steps of;

detecting a second reflectance measurement;

comparing said first reflectance measurement to said second reflectance measurement;

ending the loop if said comparison is less than or equal to a predetermined cutoff value;

replacing the value of said first reflectance measurement with the value of said second reflectance measurement; and

repeating the loop.

27. A method according to claim 26 wherein the step of determining the presence or concentration of said target comprises calculating the concentration using said second reflectance measurement.

28. A method of determining the concentration of a target in a sample comprising the steps of:

a. detecting a dry strip reading from the reagent test strip of claim 1;

b. running a preprogrammed threshold loop instruction;

c. detecting a first reflectance measurement;

d. running a preprogrammed kinetic loop instruction;

e. running a consecutive comparison instruction;

e. determining the presence or concentration of the target;

wherein said threshold loop instruction comprises the steps of;

detecting a threshold comparison measurement;

comparing said threshold comparison measurement to a predetermined threshold value;

repeating the loop until said threshold comparison measurement is less than or equal to said predetermined threshold value;

further wherein a fluid suspected of including a target is applied during the step of running said threshold loop instruction;

further wherein said kinetic loop instruction comprises the steps of;

detecting a second reflectance measurement;

comparing said first reflectance measurement to said second reflectance measurement;

ending the loop if said comparison is less than or equal to a predetermined cutoff value;

replacing the value of said first reflectance measurement with the value of said second reflectance measurement; and

repeating the loop;

further wherein said consecutive comparison instruction comprises the steps of;

detecting a third reflectance measurement;

comparing said second reflectance measurement to said third reflectance measurement;

ending the consecutive comparison instruction if said comparison is less than or equal to said predetermined cutoff value;

replacing the value of said first reflectance measurement with the value of said third reflectance measurement; and

returning to the step of running a kinetic loop instruction.

29. A method of determining the concentration of a target in a sample comprising the steps of:

a. detecting a dry strip reading from the reagent test strip of claim 1;

b. running a preprogrammed threshold loop instruction;

c. beginning a predetermined time point such that a fixed time point may be reached;

d. detecting a first reflectance measurement;

e. running a preprogrammed kinetic loop instruction;

f. determining the presence or concentration of the target;

wherein said threshold loop instruction comprises the steps of;

detecting a threshold comparison measurement;

comparing said threshold comparison measurement to a predetermined threshold value;

repeating the loop until said threshold comparison measurement is less than or equal to said predetermined threshold value;

further wherein a fluid suspected of including a target is applied during the step of running said threshold loop instruction;

further wherein said kinetic loop instruction comprises the steps of;

detecting a second reflectance measurement;

comparing said first reflectance measurement to said second reflectance measurement;

ending the loop if said second reflectance measurement is less than or equal to a predetermined cutoff value or if said fixed time point is reached;

replacing the value of the first reflectance measurement with the value of the second reflectance measurement; and

repeating the loop.

30. A method of determining the concentration of a target in a sample comprising the steps of:

a. detecting a dry strip reading from the reagent test strip of claim 1;

b. running a preprogrammed threshold loop instruction;

c. beginning a predetermined time point such that a fixed time point may be reached;

d. detecting a first reflectance measurement;

e. running a preprogrammed kinetic loop instruction;

f. running a consecutive comparison instruction;

f. determining the presence or concentration of the target;

wherein said threshold loop instruction comprises the steps of;

detecting a threshold comparison measurement;

comparing said threshold comparison measurement to a predetermined threshold value;

repeating the loop until said threshold comparison measurement is less than or equal to said predetermined threshold value;

further wherein a fluid suspected of including a target is applied during the step of running said threshold loop instruction;

further wherein said kinetic loop instruction comprises the steps of;

detecting a second reflectance measurement;

comparing said first reflectance measurement to said second reflectance measurement;

ending the loop if said second reflectance measurement is less than or equal to a predetermined cutoff value or if said fixed time point is reached;

replacing the value of the first reflectance measurement with the value of the second reflectance measurement; and

repeating the loop.

further wherein said consecutive comparison instruction comprises the steps of;

detecting a third reflectance measurement;

comparing said second reflectance measurement to said third reflectance measurement;

ending the consecutive comparison instruction if said comparison is less than or equal to said predetermined cutoff value or if said fixed time point is reached;

replacing the value of said first reflectance measurement with the value of said third reflectance measurement; and

returning to the step of running a kinetic loop instruction.

31. A method of determining the concentration of a target in a sample comprising the steps of:

a. detecting a dry strip reading from the reagent test strip of claim 1;

b. running a preprogrammed threshold loop instruction;

c. beginning a predetermined time point such that a fixed time point may be reached;

d. determining the presence or concentration of a target in a sample when said fixed time point is reached;

wherein said threshold loop instruction comprises;

detecting a threshold comparison value;

comparing said threshold comparison value to a predetermined threshold value;

repeating the loop until said threshold comparison value is less than or equal to said predetermined threshold value; and

further wherein a fluid suspected of including a target is applied during the step of running said threshold loop instruction.

23. A reagent test strip comprising:

a. a top support layer comprising a sample aperture;

b. a membrane comprising a reagent system for indicating the concentration of a target;

c. at least one spacer able to reduce contact between said membrane and a surface; and

wherein said membrane is positioned below said top support layer and in substantial alignment or approximate alignment with said sample aperture.