Immunochromatographic assay with improved colored latex

• US 5,266,497 A
• Filed: 08/30/1991
• Issued: 11/30/1993
• Est. Priority Date: 08/31/1990
• Status: Expired due to Term

• Current Assignee: JSR Corporation, Rohto Pharmaceutical Company Limited
• Original Assignee: Japan Synthetic Rubber Co., Ltd., Rohto Pharmaceutical Company Limited
• Inventors: Kinoshita, Masahiko, Nishihara, Toru, Kouda, Yoshiko, Imai, Senzo
• Primary Examiner(s): Nucker, Christine M.
• Assistant Examiner(s): PRESTON, DAVID
• Application Number:
  US07/753,231
• Time in Patent Office: 823 Days
• Field of Search: 436/518, 436/524, 436/514, 436/501, 436/533, 436/518, 436/524, 436/514, 436/528, 436/531, 436/534, 435/7.92, 435/7.93, 435/7.94, 435/7.95, 435/805, 435/970, 435/969
• US Class Current: 436/514
• CPC Class Codes: G01N 33/558 using diffusion or migration of antigen or antibody immunochromatographic test strips G01N33/54387
• CPC Class Codes: G01N 33/585 with a particulate label, e.g. coloured latex
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/969 Multiple layering of reactants
• CPC Class Codes: Y10S 435/97 Test strip or test slide

Abstract

An immunochromatography which comprises chromatographically moving a sample together with or followed by labelling fine particles in a chromatographic medium having at least one reaction site having immobilized thereat a reagent bindable to a substance to be detected in a sample, to contact the above sample and the labelling fine particles with the above reaction site, and detecting the substance to be detected by use of the phenomenon that when the above substance to be detected is present in the sample the labelling fine particles are specifically bound to the above immobilized reagent via the substance to be detected at the above reaction site, to thereby capture the substance to be detected on the chromatographic medium, characterized in that the above labelling fine particles are sensitized dyed particles obtained by sensitizing, with a material bindable to the above substance to be detected, labelling dyed particles obtained by dyeing latex particles of a synthetic high polymer, said labelling dyed particles having a surface negative charge of 0.01 to 0.5 meq/g. Said immunochromatography enables excellent visual judgement and detection of a substance to be detected which is present in a sample at a high sensitivity, and can be suitably used for the diagnosis of pregnancy.

Claims

1. An immunochromatography method which comprises the steps of:

(a) chromatographically moving a sample together with, or followed by, labelling fine particles in a chromatographic medium having at least one reaction site having immobilized thereat a reagent bindable to a substance to be detected in a sample;

(b) contacting the above sample and the labelling fine particles with the above reaction site;

(c) detecting the substance to be detected by use of the phenomenon that when said substance to be detected is present in the sample the labelling fine particles are specifically bound to said immobilized reagent via the substance to be detected at said reaction site, to thereby capture the substance to be detected on the chromatographic medium, wherein said labelling fine particles are sensitized dyed particles obtained by sensitizing said labelling fine particles with a material bindable to said substance to be detected, wherein said labelling dyed particles are obtained by dyeing latex particles of a synthetic high polymer having a functional group selected from the group consisting of --COO^- and --SO₄^- wherein said particles have a particle diameter of 0.05-0.5 μ

m, the CV value (standard deviation/average) of said diameter being 10% or less, said labelling dyed particles having a surface negative charge of about 0.01 to about 0.3 meq/g, the chroma of said dyed particles as expressed according to Munsell'"'"'s color system being such that when the dyed particles are dried, the value L* is 40 or less, the chroma C* (C*=√

a^*2 +b^*2) is 45 more and the hue angle H°

is 240°

to 60°

.

2. An immunochromatography method according to claim 1, wherein the surface negative charge of the labelling dyed particles is 0.01-0.2 meq/g.

3. An immunochromatography method according to claim 1, wherein the latex particles have particle diameters in the range of 0.05-0.3 μ

m.

4. An immunochromatography method according to claim 1, wherein the dyeing is carried out by the use of an oil-soluble dye having a solubility in toluene of at least 1 g/100 mg.

5. An immunochromatography method according to claim 1, wherein the labelling dyed particles are prepared by dyeing latex particles in water with an emulsion of an oil-soluble dye in an oily organic solvent.

6. An immunochromatography method according to claim 4, wherein the oil-soluble dye is an azo type dye or a quinone type dye.

7. An immunochromatography method according to claim 1, wherein the synthetic high polymer is a styrene-acrylic acid copolymer, a styrene-methacrylic acid copolymer or a styrene-itaconic acid copolymer.

8. The immunochromatography method according to claim 1, wherein the chromatographic medium is a filter comprises an inorganic fiber.