Non-instrumented cholesterol assay

• US 5,340,539 A
• Filed: 10/08/1992
• Issued: 08/23/1994
• Est. Priority Date: 03/16/1989
• Status: Expired due to Term

• Current Assignee: Chemtrak
• Original Assignee: Chemtrak
• Inventors: Jeong, Henry J., Allen, Michael P.
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): EDWARDS, LAURA ESTELLE
• Application Number:
  US07/958,519
• Time in Patent Office: 684 Days
• Field of Search: 422/56, 422/57, 422/58, 436/169, 436/170, 435/970, 435/11, 435/28
• US Class Current: 422/421
• CPC Class Codes: C12Q 1/00 Measuring or testing processes involving enzymes, nucleic acids or microorganisms measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34; Compositions therefor; Processes of preparing such compositions
• CPC Class Codes: C12Q 1/26 involving oxidoreductase
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: C12Q 1/60 involving cholesterol
• CPC Class Codes: G01N 33/523 the element being adapted for a specific analyte
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: G01N 33/92 involving lipids, e.g. cholesterol , lipoproteins, or their receptors steroid hormones G01N33/743
• CPC Class Codes: Y10S 435/97 Test strip or test slide

Abstract

An assay for determining the cholesterol level in a sample involving a bibulous strip comprising a transfer region for transporting a transport medium from a transport medium source, a sample receiving region in fluid communication with the transfer region, and a measurement region in fluid communication with the sample receiving region, and a detectable signal reagent system comprising a catalytic agent or enzyme, unbound conversion reagent and bound reagent wherein the conversion reagent reacts with cholesterol to form an intermediate product and wherein the bound reagent reacts with the intermediate product in the presence of the catalytic agent or enzyme to produce a detectable border, in which the conversion reagent is placed in the transfer region of the strip or in a region of the strip between the sample receiving and measurement regions and the signal reagent is non-diffusively bound to the strip in the measurement region, and which upon contact with the sample and the transport medium results in the production of a detectable border in the measurement region which is related to the level of cholesterol in the sample.

Claims

1. A device for determining the cholesterol or cholesterol ester level in a sample, said device comprising:

(a) a first bibulous strip comprising a transfer region for transporting a transport medium from a transport medium source,

(b) a bibulous member comprising a sample receiving region;

(c) a second bibulous strip comprising a conversion reagent region,

(d) a third bibulous strip comprising a measurement region;

wherein said sample receiving region is in fluid communication with said transfer, conversion, and measurement regions, or can be moved into fluid transfer relationship with said transfer, conversion, and measurement regions;

such that when said sample receiving region is moved into fluid transfer relationship with said transfer, conversion, and measurement regions, said device further comprises;

(e) means for moving said sample receiving region from a first site for receiving said sample to fluid transfer relationship with said transfer, conversion, and measurement regions;

(f) conversion reagent capable of reacting with cholesterol and cholesterol ester to form an intermediate product, said conversion reagent present on said second strip upstream from said measurement region, any additional conversion reagent being present on the sample receiving region, on the transfer region, or in said transport medium, such that movement of the transport medium through said second strip will bring the conversion reagent and cholesterol together to react to form said intermediate product; and

(g) non-diffusively bound reagent in the measurement region which reacts in the presence of said intermediate product to produce a detectable border related to the cholesterol or cholesterol ester level in said sample.

2. The device of claim 1, further comprising means for providing a catalytic agent capable of causing a reaction with said intermediate product to result in a reaction with said non-diffusively bound reagent, such that when said means for providing a catalytic agent provides said catalytic agent in said measurement region said catalytic agent is non-diffusively bound throughout said measurement region.

3. The device of claim 1, further comprising horseradish peroxidase in the transport medium, on the transfer region or on the region between the sample receiving and measurement region.

4. The device of claim 2, wherein said conversion reagent comprises cholesterol esterase and cholesterol oxidase and said intermediate product is hydrogen peroxide.

5. The device of claim 4, wherein said catalytic agent is horseradish peroxidase.

6. The device of claim 2, wherein said device further comprises a capture region on said sample region or upstream from said measurement region, which capture region diminishes a predetermined portion of reaction of said bound reagent in said measurement region.

7. The device of claim 2, wherein said non-diffusively bound reagent is a leuco dye.

8. The device of claim 2, wherein said device comprises a transport solution container.

9. The device of claim 8, wherein said transport solution comprises at least one or more of cholesterol esterase, cholesterol oxidase or peroxidase.

10. A device for determining the cholesterol or cholesterol ester level in a sample, said device comprising:

(a) a first bibulous strip comprising a transfer region for transporting a transport medium from a transport medium source;

(b) a bibulous member comprising a sample receiving region;

(c) a second bibulous strip comprising a conversion region;

(d) a third bibulous strip comprising a measurement region;

wherein said sample receiving region is in fluid communication with said transfer, conversion, and measurement regions, or can be moved into fluid transfer relationship with said transfer, conversion, and measurement regions;

such that when said sample receiving region is moved into fluid transfer relationship with said transfer, conversion, and measurement regions, said device further comprises;

(e) means for moving said sample receiving region from a first site for receiving said sample to fluid transfer relationship with said transfer, conversion, and measurement regions;

(f) conversion reagent comprising cholesterol esterase and cholesterol oxidase capable of reacting with cholesterol and cholesterol ester to form hydrogen peroxide, said conversion reagent being present on said second strip, said second strip being present in a region between the sample receiving and measurement regions such that movement of the transport medium through said second strip will bring the conversion reagent and cholesterol together to react to form an intermediate product;

(g) non-diffusively bound reagent in the measurement region which reacts in the presence of said intermediate product to produce a detectable border related to the cholesterol or cholesterol ester level in said sample; and

(h) means for providing a peroxidase agent capable of causing a reaction with said hydrogen peroxide, such that when said means for providing a peroxidase provides said peroxidase in said measurement region said peroxidase is non-diffusively bound throughout said measurement region.

11. The device of claim 10, wherein said intermediate product is hydrogen peroxide.

12. The device of claim 11, wherein said peroxidase is horseradish peroxidase.

13. The device of claim 10, wherein said device further comprises a capture region on said sample region or upstream from said measurement region, which capture region diminishes a predetermined portion of reaction of said bound reagent in said measurement region.

14. The device of claim 13, wherein said capture region is downstream from said sample receiving region and comprises a reactant capable of reacting with hydrogen peroxide.

15. The device of claim 10, wherein said non-diffusively bound reagent is a leuco dye.

16. The device of claim 10, wherein said sample receiving region is out of fluid transfer relationship with said transfer and measurement regions and further comprising means for moving said sample receiving region from a first site for receiving said sample to fluid transfer relationship with said transport and measurement regions.