One step immunochromatographic device and method of use

• US 5,712,172 A
• Filed: 04/18/1995
• Issued: 01/27/1998
• Est. Priority Date: 05/18/1995
• Status: Expired due to Term

• Current Assignee: Sekisui Diagnostics, LLC (Sanofi )
• Original Assignee: Wyntek Diagnostics, Inc. (Genzyme General)
• Inventors: Fan, Eugene, Huang, Ching
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US08/444,238
• Time in Patent Office: 1,015 Days
• Field of Search: 422/56-58, 435/287.1, 435/287.2, 435/287.7, 435/287.9, 435/805, 435/810, 435/967, 435/970, 435/971, 436/169, 436/510, 436/514, 436/518, 436/523, 436/525, 436/531, 436/534, 436/805, 436/810, 436/65, 436/814, 436/818
• US Class Current: 436/518
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/967 Standards, controls, materials, e.g. validation studies, buffer systems
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 435/971 Capture of complex after antigen-antibody reaction
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/814 Pregnancy
• CPC Class Codes: Y10S 436/818 Human chorionic gonadotropin

Abstract

This invention relates to a lateral flow immunochromatographic assay device without a plastic housing. The assay includes a sample receiving region, analyte detection region and end region all made of porous material and capable of lateral flow. The analyte detection region includes labeling reagents, a capture reagent and a control reagent. The back of the porous material is laminated with a semi-rigid material with adequate mechanical strength. The top is partially covered with a thin plastic material so as to leave a portion of the sample receiving region exposed for sample application.

Claims

1. An immunochromatographic assay device for detection of the presence or absence of an analyte in a liquid sample, wherein said immunochromatographic assay device consists of:

(a) a separate sample receiving region comprising a porous material which conducts lateral flow of a liquid sample, in lateral flow contact with

(b) a separate analyte detection region comprising a porous material which conducts lateral flow of said liquid sample, wherein said analyte detection region comprises a first and a second mobile labeling reagent at a discrete labeling situs, an immobile capture reagent at a discrete capture situs, and an immobile control reagent at a discrete control situs, wherein said first labeling reagent is capable of forming a complex with said analyte;

said capture reagent is capable of binding said analyte-labeling reagent complex and said control reagent is capable of binding said second labeling reagent, wherein said analyte detection region is in lateral flow contact with

(c) a separate end flow region comprising a porous material which conducts lateral flow of said liquid sample and capable of absorbing excess liquid sample,

wherein said liquid sample laterally flows from said sample receiving region towards said end flow region, and mixes with said first and second labeling reagents to move said first and second labeling reagents towards said end flow region, wherein the back of said porous materials are laminated with a semi-rigid material of at least 0.005 inches thick to produce a device with adequate mechanical strength, and the front of said porous materials partially covered with a plastic material so as to leave a portion of said sample receiving region exposed for sample application and allow the discrete capture situs and said discrete control situs to be visible.

2. The device of claim 1 wherein said capture reagent is antibody or an immunologically reactive fragment thereof specific for said analyte.

3. The device of claim 1 wherein said first labeling reagent is an antibody or an immunologically reactive fragment thereof specific for said analyte, and said second labeling reagent is not immunologically reactive to said analyte, wherein said first labeling reagent is attached to a first labeling particle and said second labeling reagent is attached to a second labeling particle.

4. The device of claim 1 wherein said porous material is nitrocellulose, nitrocellulose blends with polyester, cellulose, paper or nylon.

5. The device of claim 1 wherein said semi-rigid material is vinyl.

6. The device of claim 1 wherein said plastic material is mylar.

7. The device of claim 1 wherein said control reagent is antibody or an immunologically reactive fragment specific for said second labeling reagent.

8. The device of claim 3 wherein said particles are colored latex, dye sols, colloidal gold or liposomes containing dye.

12. A method to determine the presence or absence of analyte in a sample, which method comprises applying said sample to the sample receiving region of the device of claims 1, 9, 10 or 11, so as to permit said sample to flow through the analyte detection region and into the end flow region, and

detecting the presence or absence of analyte in the analyte detection region at the discrete capture reagent situs containing the capture reagent capable of binding the analyte-labeling reagent complex or the discrete control situs containing the control reagent capable of binding the labeling reagent.

13. The method of claim 12 wherein said analyte is human chorionic gonadotropin (hCG).

14. The method of claim 13 wherein said first labeling reagent is a monoclonal antibody immunoreactive with the β

-epitope of hCG conjugated to blue latex and said second labeling reagent is BSA conjugated to red latex.

15. The method of claim 14 wherein said capture reagent is a monoclonal antibody immunoreactive with the α

-epitope of hCG.

16. The method of claim 15 wherein said control reagent is a monoclonal antibody immunoreactive with the BSA-conjugated red latex.

17. The method of claim 12 wherein said detection occurs within one minute of applying the sample.

9. An immunochromatographic assay device for detection of the presence or absence of an analyte in a liquid sample, wherein said immunochromatographic assay device consists of:

(a) a separate sample receiving region comprising a porous material which conducts lateral flow of a liquid sample, wherein said sample receiving region comprises a first and second mobile labeling reagent at a discrete labeling situs, wherein said first labeling reagent is capable of forming a complex with said analyte, wherein said sample receiving region is in lateral flow contact with

(b) a separate analyte detection region comprising a porous material which conducts lateral flow of said liquid sample, wherein said material region comprises an immobile capture reagent at a discrete capture situs, and an immobile control reagent at a discrete control situs, wherein said capture reagent is capable of binding said analyte-labeling reagent complex and said control reagent is capable of binding said second labeling reagent, wherein said analyte detection region is in lateral flow contact with

(c) a separate end flow region comprising a porous material which conducts lateral flow of said liquid sample and capable of absorbing excess liquid sample,

wherein said liquid sample laterally flows from said sample receiving region towards said end flow region, and mixes with said first and second labeling reagents to move said first and second labeling reagents towards said end flow region, wherein the back of said porous materials are laminated with a semi-rigid material of at least 0.005 inches thick to produce a device with adequate mechanical strength, and the front of said porous materials partially covered with a plastic material so as to leave a portion of said sample receiving region exposed for sample application and allow the discrete capture situs and said discrete control situs to be visible.

10. An immunochromatographic assay device for detection of the presence or absence of an analyte in a liquid sample, wherein said immunochromatographic assay device consists of:

(a) a separate sample receiving region comprising a porous material which conducts lateral flow of a liquid sample, in lateral flow contact with

(b) a labeling reagent region comprising a first and second mobile labeling reagent, wherein said first labeling reagent is capable of binding said analyte-labeling reagent complex, wherein said labeling reagent region is in lateral flow contact with

(c) a separate analyte detection region comprising a porous material which conducts lateral flow of said liquid sample, wherein said separate analyte region comprises an immobile capture reagent at a discrete capture situs, and an immobile control reagent at a discrete control situs, wherein said capture reagent is capable of binding said analyte-labeling reagent complex and said control reagent is capable of binding said second labeling reagent, wherein said analyte detection region is in lateral flow contact with

(c) a separate end flow region comprising a porous material which conducts lateral flow of said liquid sample and capable of absorbing excess liquid sample,

wherein said liquid sample laterally flows from said sample receiving region towards said end flow region, and mixes with said first and second labeling reagents to move said first and second labeling reagents towards said end flow region, wherein the back of said porous materials are laminated with a semi-rigid material of at least 0.005 inches thick to produce a device with adequate mechanical strength, and the front of said porous materials partially covered with a plastic material so as to leave a portion of said sample receiving region exposed for sample application and allow the discrete capture situs and said discrete control situs to be visible.

11. An immunochromatographic assay device for detection of the presence or absence of an analyte in a liquid sample, wherein said immunochromatographic assay device consists of;

(a) a separate sample receiving region comprising a porous material which conducts lateral flow of a liquid sample, in lateral flow contact with

(b) a labeling reagent region comprising a first and second mobile labeling reagent, wherein said first labeling reagent is capable of binding said analyte-labeling reagent complex, wherein said labeling reagent region is in lateral flow contact with

(c) a separate analyte detection region comprising a porous material which conducts lateral flow of said liquid sample, wherein said separate analyte region comprises an immobile capture reagent at a discrete capture situs, and an immobile control reagent at a discrete control situs, wherein said capture reagent is capable of binding said analyte-labeling reagent complex and said control reagent is capable of binding said second labeling reagent, wherein said analyte detection region is in lateral flow contact with

(c) a separate end flow region comprising a porous material which conducts lateral flow of said liquid sample and capable of absorbing excess liquid sample,

wherein said liquid sample laterally flows from said sample receiving region towards said end flow region, and mixes with said first and second labeling reagents to move said first and second labeling reagents towards said end flow region, wherein the back of said porous materials are laminated with a semi-rigid material of at least 0.005 inches thick to produce a device with adequate mechanical strength, and the front of said porous material is partially covered with a plastic material such that the portion covering the sample receiving region has holes or openings to expose the sample to the sample receiving region and allow the discrete capture situs and said discrete control situs to be viewed.