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

Patent metadata

Field	Value
Publication number	US-10422798-B2
Application number	US-201415536621-A
Country	US
Kind code	B2
Filing date	Dec 16, 2014
Priority date	Dec 16, 2014
Publication date	Sep 24, 2019
Grant date	Sep 24, 2019

Abstract

A problem of the present invention is to provide an immunochromatographic test strip avoiding agglutination of colloidal gold while red blood cells in whole blood are agglutinated and then separated and removed in the case of using polybrene as a hemagglutinating agent and the colloidal gold conjugates as a detection reagent, and to provide immunochromatography using the test strip. To solve the problem, the present inventors reviewed the composition of the existing reagent itself from a completely different viewpoint rather than the selection of type or amount of polyanions, and as a result of extensive study on each element, the inventors surprisingly found that agglutination of colloidal gold may be suppressed by using a particular additive without 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 hexadimethrine bromide is contained in the sample supply portion, and
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 downstream from the conjugate-retaining part; and
(2) an additive in a dry state, wherein the additive is contained in the sample-supply portion or the conjugate-retaining part of the spreading portion so as to be capable of contacting with hexadimethrine bromide that is provided through the sample-supply portion,
wherein the additive is one or more compounds selected from the group consisting of (A) a salt of a divalent metal, (B) a salt of sulfuric acid with a metal or onium, (C) a salt of sulfurous acid with a metal, and (E) a polybasic amino acid,
wherein a polyanion as a neutralizer for neutralizing a cation of the hexadimethrine bromide is not included on the test strip.

2. The test strip of claim 1 , wherein the additive is contained in the conjugate-retaining part.

3. A detection method of using immunochromatography, comprising the steps of:
(A) supplying a sample to a sample-supply portion of a test strip, said test strip comprising a membrane, said membrane comprising a porous body, which comprises the sample-supply portion and a spreading portion,
wherein the sample-supply portion is upstream from the spreading portion,
wherein hexadimethrine bromide is contained in the sample-supply portion,
wherein a conjugate comprising an anti-analyte antibody immobilized on 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 downstream from the conjugate-retaining part,
wherein a polyanion as a neutralizer for neutralizing a cation of the hexadimethrine bromide is not included on the test strip,
wherein an additive is contained in the sample-supply portion or the conjugate-retaining part of the spreading portion in a dry state, and
wherein the additive is one or more compounds selected from the group consisting of (A) a salt of a divalent metal, (B) a salt of sulfuric acid with a metal or onium, (C) a salt of sulfurous acid with a metal, and (E) a polybasic amino acid;
(B) agglutinating red blood cells in the sample through contact between hexadimethrine bromide and the sample in the sample-supply portion;
(C) separating and removing aggregates obtained in step (B) from the sample;
(D) bringing a sample component obtained in step (C), after separation and removal of the aggregates, into contact with the colloidal gold-containing conjugate, step (D) being performed in the presence of the additive; and
(E) detecting a complex of the analyte in the sample component and the conjugate obtained in step (D) with the antibody immobilized in the detecting portion.

4. The detection method of claim 3 , wherein the additive is contained in the conjugate-retaining part.

5. The detection method of claim 3 , wherein the sample is a heparin-containing sample or an EDTA-containing sample.

Assignees

• Sekisui Medical Co Ltd

Inventors

• Kohno Keigo
• Yoshida Mayumi
• Yamamoto Mitsuaki
• Yokokawa Shinya
• Asami Marie

Classifications

• G01N33/543

  with an insoluble carrier for immobilising immunochemicals · 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

• G01N33/54386

  Analytical elements · CPC title

• G01N33/558

  using diffusion or migration of antigen or antibody {(immunochromatographic test strips G01N33/54387)} · CPC title

• G01N33/54388 Primary

  based on lateral flow · CPC title