Direct measuring assay strip and method of use thereof

• US 4,999,287 A
• Filed: 05/19/1988
• Issued: 03/12/1991
• Est. Priority Date: 05/19/1988
• Status: Expired due to Term

• Current Assignee: Chemtrak
• Original Assignee: Chemtrak
• Inventors: Allen, Michael P., Shibuya, Robert B.
• Primary Examiner(s): Hill, Jr., Robert J.
• Application Number:
  US07/195,881
• Time in Patent Office: 1,027 Days
• Field of Search: 436/169, 436/805, 436/817, 422/56, 422/58, 422/60, 422/61, 435/11, 435/14, 435/805, 435/810, 435/25, 435/28
• US Class Current: 435/11
• 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 33/525 Multi-layer analytical elements
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/81 Packaged device or kit

Abstract

Methods and compositions are provided for measuring an analyte without instrumentation. The device employs a stripstick which may include as its components, a discontinuous flow path, which is made continuous after transfer of the sample, means for automatically metering the volume of a sample, and means for providing a sharply delineated color front. The method finds particular use where a limited amount of substrate is provided for an enzyme on a sample pad.

Claims

1. A dipstick measuring device comprising:

in a direction of eluent flow,

a first bibulous bridging strip extending from an immersion end of said device to a sample pad site;

a sample pad at said sample pad site;

a second bibulous bridging strip extending from said sample pad site to a measurement region fluid receiving site; and

an extended bibulous measurement strip in fluid communication with said second bridging strip and impregnated with a first member of a signal producing system, which upon reaction with a second member of said signal producing system produces a detectable signal defining a boundary on said measurement strip; and

a removable inert protective barrier separating said sample pad from fluid communication with said first and second bridging strips and bringing said sample paid into fluid communication with said first and second bridging strips when removed.

2. A device according to claim 1, wherein said first and second bridging strips overlap said sample pad without contact between said strips.

3. A device according to claim 1, wherein said first and second bridging strips, said sample pad and said bibulous measurement strip are bound to and supported by a supporting film.

4. A device according to claim 1, wherein said signal producing system comprises at least one enzyme impregnated in said sample pad.

5. A device according to claim 4, wherein said enzyme is an oxidase.

20. A kit comprising a device according to claim 1, members of said signal producing system, and a sample metering device comprising a wicking mesh backed with an absorbant layer.

21. A kit according to claim 20, wherein said signal producing system comprises horseradish peroxidase.

6. A dipstick measuring device comprising:

in a direction of eluent flow,

a first bibulous bridging strip extending from an immersion end of said device to a sample pad site;

a sample pad at said sample pad site;

a second bibulous bridging strip extending from said sample pad site to a measurement region fluid receiving site; and

an extended bibulous measurement strip in fluid communication with said second bridging strip and impregnated with a first member of a signal producing system, which upon reaction with a second member of said signal producing system produces a detectable signal defining a boundary on said measurement strip; and

a removable inert protective barrier separating said sample pad from fluid communication with said first and second bridging strips and bringing said sample pad into fluid communication with said first and second bridging strips when removed said sample pad and said bibulous measurement strip are bound to and supported by a supporting film, and said supporting film has a sample port beneath said sample pad.

7. A dipstick measuring device comprising:

in a direction of eluent flow,

a first bibulous bridging strip extending from an immersion end of said device to a sample pad site;

a sample pad at said sample pad site;

a second bibulous bridging strip extending from said sample pad site to a measurement region fluid receiving site; and

an extended bibulous measurement strip in fluid communication with said second bridging strip and impregnated with a first member of a signal producing system, which upon reaction with a second member of said signal producing system produces a detectable signal defining a boundary on said measurement strip; and

in addition

a removable inert protective barrier separating said sample pad from fluid communication with said first and second bridging strips and bringing said sample pad into fluid communication with said first and second bridging strips when removed; and

a spanning bibulous strip extending from said immersion end to said measurement strip in fluid communication with said measurement strip; and

wherein said second bridging strip has a region narrower than said spanning strip.

8. A device according to claim 7, wherein said spanning bibulous strip intervenes between and provides fluid communication between said second bridging strip and said measurement strip.

9. A device according to claim 7, wherein said narrower region of said second bridging strip is of from about 20 to 80% of the width of said spanning strip.

10. A dipstick measuring device comprising:

in a direction of eluent flow,

a first bibulous bridging strip extending from an immersion end of said device to a sample pad site;

a sample pad at said sample pad site;

a second bibulous bridging strip extending from said sample pad site to a measurement region fluid receiving site; and

an extended bibulous measurement strip in fluid communication with said second bridging strip and impregnated with a first member of a signal producing system, which upon reaction with a second member of said signal producing system produces a detectable signal defining a boundary on said measurement strip; and

a removable metering device comprising a porous non-wetting member in fluid receiving contact with said sample pad, an absorbant layer backing for receiving fluid from said sample pad, covered with an inert film providing an inert protective barrier separating said sample pad from fluid communication with said first and second bridging strips, said removable metering device bringing said sample pad into fluid communication with said first and second bridging strips when removed.

11. A dipstick measuring device comprising:

in a direction of eluent flow;

a first bibulous bridging strip extending from an immersion end of said device to a sample pad site;

a sample at said sample pad site;

a second bibulous bridging strip extending from said sample pad site to a measurement region fluid receiving site;

an extended bibulous measurement strip in fluid communication with said second bridging strip and impregnated with a first member of a signal producing system, which upon reaction with a second member of said signal producing system, produces a detectable signal defining a boundary on said measurement strip; and

a removable metering device comprising a porous;

non-wetting member in fluid receiving contact with said sample pad, said porous non-wetting member consisting essentially of a plastic screen of about 250 μ

m to about 1 mm and a thickness in the range of about 150 to 600 μ

m, an absorbent layer backing for receiving fluid from said sample pad, covered with an inert film providing an inert protective barrier separating said sample pad from fluid communication with said first and second bridging strips, said removable metering device bringing said sample pad into fluid communication with said first and second bridging strips when removed.

12. A dipstick measuring device comprising:

in the direction of eluent flow,

a first bibulous bridging strip extending from an immersion end of said device to a sample pad site;

a sample pad at said sample pad site;

a second bibulous bridging strip extending from said sample pad site to a measurement region fluid receiving site; and

an extended bibulous measurement strip in fluid communication with said second bridging strip and impregnated with a first member of a signal producing system, which upon reaction with a second member of said signal producing system produces a detectable signal defining a boundary on said measurement strip; and

in addition

a spanning bibulous strip extending from said immersion end to said measurement strip in fluid communication with said measurement strip;

wherein said second bridging strip has a region narrower than said spanning strip;

a removable metering device comprising a porous non-wetting member in fluid receiving contact with said sample pad, an absorbant layer backing for receiving fluid from said sample pad, covered with an inert film providing an inert protective barrier separating said sample pad from fluid communication with said first and second bridging strips, and removable metering device bringing said sample pad into fluid communication with said first and second bridging strips when removed.

13. A method for measuring the presence of an analyte in a sample, employing a device comprising a first bridging bibulous member for transferring eluent from a fluid source to a sample pad, a second bridging member for transferring eluent fluid from said sample pad to a measuring extended bibulous strip impregnated with a second member of a signal producing system, said method comprising:

impregnating a sample pad with an assay medium resulting in the presence of a first member of a signal producing system in said sample pad in an amount related to the amount of analyte in said sample, while preventing transport of said assay medium to said bridging bibulous members;

directing an eluent stream from an eluent source to said sample pad to transfer said first member from said pad to said measuring strip through said second bridging member;

directing said first member containing eluent through said measuring strip, whereby said first and second members of said signal producing system and any additional members of said signal producing system react to produce a detectable boundary on said measuring strip, wherein the extent of said is related to the amount of analyte in said sample.

14. A method for measuring the presence of an analyte in a sample, employing a device comprising:

a first bridging bibulous member for transferring eluent from a fluid source to a sample pad;

a second bridging member for transferring eluent fluid from said sample pad to a measuring extended bibulous strip impregnated with a second member of a signal-producing system, said method comprising;

impregnating a sample pad with an assay medium, resulting in the presence of a first member of a signal-producing system in said sample pad in an amount related to the amount of analyte in said sample, while preventing transport of said assay medium to said bridging bibulous members;

directing an eluent stream from an eluent source to said sample pad to transfer said first member from said pad to said measuring strip through said second bridging member;

directing said first member containing eluent through said measuring strip;

directing a diverted stream directly from said eluent source to said measuring member, wherein said diverted stream extends the width of said measuring member and said first member containing eluent stream is narrower than the width of said measuring member and is directed to the middle of said measuring member;

whereby said first and second members of said signal producing system and any additional members of said signal producing system react to produce a detectable boundary on said measuring strip, when the extent of said boundary is related to the amount of analyte in said sample.

15. A method according to claim 14, further comprising;

impregnating said sample pad with a greater than saturating amount of said assay medium; and

wicking excess assay medium through a plastic mesh into an absorbant layer; and

removing said plastic mesh and absorbant layer from said sample pad prior directing said eluent stream to said sample pad,

whereby said assay medium is measured in a reproducible manner.

16. A method according to claim 14, whereby said signal producing system comprises an oxidase covalently immobilized in said sample pad, a dialkylaniline covalently immobilized in said measuring strip, a hydrazine impregnated in said measuring strip and a peroxidase in said eluent.

17. A method according to claim 16, wherein said oxidase is glucose oxidase and said analyte is glucose.

18. A method according to claim 16, wherein said oxidase is alcohol oxidase and said analyte is an alcohol.

19. A method according to claim 16, wherein said oxidase is cholesterol oxidase and said analyte is cholesterol and said signal producing system further comprises cholesterol esterase impregnated in said sample pad.