Immunochromatographic test strip and detection method using immunochromatography for detecting object in red blood cell-containing sample

Patent metadata

Field	Value
Publication number	US-9885707-B2
Application number	US-201314389180-A
Country	US
Kind code	B2
Filing date	Apr 1, 2013
Priority date	Mar 30, 2012
Publication date	Feb 6, 2018
Grant date	Feb 6, 2018

Abstract

A problem to be solved by the present invention is to provide an immunochromatographic test strip and a detection method using immunochromatography avoiding aggregation of colloidal gold conjugates while red blood cells in whole blood are agglutinated and then separated and removed in the case of using polybrene as a blood-agglutinating agent and the colloidal gold conjugates as a detection reagent. To solve the problem, the present inventers reviewed a past reagent configuration itself from a completely different viewpoint rather than selecting type and amount of polyanions and, as a result of extensive study on each element, the inventers surprisingly found that aggregation of colloidal gold can be suppressed by using a certain buffer solution without using neutralization by polyanions.

First claim

The invention claimed is:

1. An immunochromatographic test strip, comprising:
(1) a membrane comprising a porous body, which comprises a sample-supply portion and a spreading portion,
wherein the sample-supply portion is upstream from the spreading portion,
wherein the sample-supply portion contains hexadimethrine bromide,
wherein a conjugate of an anti-analyte antibody labeled with colloidal gold is retained in a dissoluble manner in a conjugate-retaining part of the spreading portion, and an antibody is immobilized in a detecting part of the spreading portion, which is from the conjugate-retaining part,
wherein the sample-supply portion is a sample pad, and
wherein the conjugate-retaining part is a conjugate pad;
(2) a buffer solution component in a dry state,
wherein the buffer solution component is contained on the porous body upstream from the conjugate-retaining part so as to be able to contact with the hexadimethrine bromide provided through the sample-supply portion, and
wherein the buffer solution component is selected from the group consisting of Bis-Tris, MES, and citric acid.

2. The test strip of claim 1 , wherein the sample-supply portion comprises the buffer solution component.

3. The test strip of claim 1 , wherein the test strip does not comprise a polyanion.

4. The test strip of claim 1 , wherein the buffer solution component is Bis-Tris.

5. The test strip of claim 1 , wherein the buffer solution component is MES.

6. The test strip of claim 1 , wherein the buffer solution component is citric acid.

7. The test strip of claim 1 , wherein the sample-supply portion comprises glass fiber.

8. The test strip of claim 1 , wherein the amount of the buffer solution component is an amount capable of suppressing the aggregation of the colloidal gold conjugate due to the hexadimethrine bromide.

9. The test strip of claim 1 , wherein the sample pad is obtained by impregnating the sample pad with 20 to 100 mmol/L Bis-Tris, MES, or citric acid buffer solution and drying.

10. The test strip of claim 9 , wherein the concentration of hexadimethrine bromide in said 20 to 100 mmol/L Bis-Tris, MES, or citric acid buffer solution is 0.5 to 4%.

11. The test strip of claim 8 , wherein the sample pad is obtained by impregnating the sample pad with 20 to 100 mmol/L Bis-Tris, MES, or citric acid buffer solution and drying.

12. The test strip of claim 11 , wherein the concentration of hexadimethrine bromide in said 20 to 100 mmol/L Bis-Tris, MES, or citric acid buffer solution is 0.5 to 4%.

13. The test strip of claim 1 , wherein the sample pad is obtained by impregnating the sample pad with a solution containing 0.5 to 4% hexadimethrine bromide.

14. The test strip of claim 1 , wherein the sample pad is obtained by impregnating the sample pad with a solution containing 20 to 100 mmol/L Bis-Tris, MES, or citric acid buffer and a solution containing 0.5 to 4% hexadimethrine bromide and drying.

15. The test strip of claim 8 , wherein the sample pad is obtained by impregnating the sample pad with a solution containing 0.5 to 4% hexadimethrine bromide.

16. The test strip of claim 8 , wherein the sample pad is obtained by impregnating the sample pad with a solution containing 20 to 100 mmol/L Bis-Tris, MES, or citric acid buffer and a solution containing 0.5 to 4% hexadimethrine bromide and drying.

17. An immunochromatographic test strip, comprising:
(1) a sample pad suitable to be supplied with a sample;
(2) a conjugate pad disposed downstream of the sample pad and retaining in a dissoluble manner a conjugate of an anti-analyte antibody labeled with colloidal gold; and
(3) an insoluble membrane disposed downstream of the conjugate pad and immobilizing an antibody which binds to a complex of the conjugate and an analyte,
wherein the sample pad comprises hexadimethrine bromide, and
wherein the sample pad and the conjugate pad comprise a buffer solution component selected from the group consisting of Bis-Tris, MES and citric acid,
wherein the buffer solution component is in a dry state.

Assignees

• Sekisui Medical Co Ltd

Inventors

• Yoshida Mayumi
• Yamamoto Mitsuaki
• Yokokawa Shinya
• Shimizu Tomo
• Kono Keigo

Classifications

• G01N33/5306 Primary

  Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding · CPC title

• G01N33/54393 Primary

  Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding · CPC title

• G01N33/54366

  Apparatus specially adapted for solid-phase testing · CPC title

• B01L2300/0825

  Test strips · CPC title

• G01N33/5005

  involving human or animal cells (immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025) · CPC title