Immunoassay method to prevent inhibition of antigen-antibody binding interactions in mucosal fluids

Patent metadata

Field	Value
Publication number	US-11719695-B2
Application number	US-201816498579-A
Country	US
Kind code	B2
Filing date	Mar 27, 2018
Priority date	Mar 27, 2017
Publication date	Aug 8, 2023
Grant date	Aug 8, 2023

Abstract

The purpose of the present invention is to: provide an agent that effectively suppresses inhibition of antigen-antibody reaction in an immunoassay using a sample containing a body fluid, in particular, a component derived from a biological mucosal membrane, such as saliva; and to suppress false positive and false negative results in the immunoassay. The present invention provides an agent for suppressing inhibition of immune reaction, characterized in that the agent comprises a compound of the following (1) or (2): (1) Sulfonic acid compound of the formula R1—SO3H or a salt thereof. (In the formula, R1 is selected from the group consisting of: a straight-chain C5-C30 alkyl group; a straight-chain C1-C30 alkyl group substituted with an aryl group having at least one straight-chain C5-C30 alkyl group; and an aryl group having at least one straight-chain C5-C30 alkyl group. These groups may include a substituent group); and (2) Quaternary ammonium ion of the formula N+—R2R3R4R5 or a salt thereof. (In the formula, R2—R5 are each independently a straight-chain C1-C30 alkyl group, or an aryl group substituted with at least one straight-chain C5-C30 alkyl group. These groups may include a substituent group); wherein the agent is capable of suppressing immune reaction inhibitory action caused by a body fluid in an immunoassay sample.

First claim

The invention claimed is:

1. An immunoassay method for preventing inhibition of antigen-antibody binding interactions by a mucosal fluid, comprising,
(a) preparing a test sample comprising a mucosal fluid from a subject,
(b) bringing the test sample into contact with a target antigen-specific antibody,
(c) adding an agent to suppress inhibition of antigen-antibody binding in the test sample, and
(d) detecting the antigen-antibody binding in the test sample using antigen-antibody reaction
wherein the agent comprises:
a sulfonic acid compound of the formula R 1 —SO 3 H, or a salt thereof, or
a quaternary ammonium ion of the formula N + —R 2 R 3 R 4 R 5 or a salt thereof,

wherein R 1 is an unsubstituted straight-chain C 5 -C 20 alkyl group, a straight-chain C 1 -C 20 alkyl group substituted with a phenyl group having a straight-chain C 5 -C 20 alkyl group, or a phenyl group substituted with a straight-chain C 5 -C 20 alkyl group, and
wherein R 2 to R 5 are each independently a straight-chain C 1 -C 20 alkyl group unsubstituted or substituted with a phenyl group, and further
wherein the agent is capable of suppressing false positive and/or false negative results of antigen-antibody binding caused by the mucosal fluid in the test sample.

2. The method according to claim 1 ,
wherein in the formula of R 1 —SO 3 H, R 1 is an unsubstituted straight-chain C 5 -C 20 alkyl group, or a phenyl group substituted with a straight-chain C 5 -C 20 alkyl group.

3. The method according to claim 1 ,
wherein in the formula of N + —R 2 R 3 R 4 R 5 , R 2 to R 5 are each independently an unsubstituted straight-chain C 1 -C 20 alkyl group, or
R 2 to R 4 are each independently an unsubstituted straight-chain C 1 -C 20 alkyl group and R 5 is a straight-chain C 1 -C 20 alkyl group substituted with a phenyl group.

4. The method according to claim 1 ,
wherein the salt of the ammonium ion is formed between the ammonium ion and a compound selected from the group consisting of hydrogen halide, nitric acid, hexafluorophosphoric acid, dihydrogen trifluoride, and perchloric acid.

5. The method according to claim 1 ,
wherein the immunoassay is selected from the group consisting of an ELISA, an immunochromatography method, a Western blot method, an immunoblot method, an immunoprecipitation method, and a latex agglutination method.

6. The method according to claim 5 ,
wherein the test sample comprises, as the mucosal fluid from the subject, at least one of saliva, sputum, a throat swab, a nasal swab, a nasal aspirate, and a keratoconjunctive swab.

7. The method according to claim 5 ,
wherein the test sample comprises an antigen or antibody from a pathogen, a pathogenic microorganism, a hormone, or a prostaglandin.

8. The method according to claim 7 ,
wherein the pathogen or the pathogenic microorganism is a pathogen or a pathogenic microorganism, selected from the group consisting of a foot-and-mouth disease virus, an influenza virus, an adenovirus, a respiratory syncytial virus, a coronavirus, a rabies virus, a Bordetella pertussis , a hemolytic streptococcus , a food poisoning bacterium, a Chlamydia trachomatis , and a mycoplasma.

9. A method for detecting a pathogen-specific antigen in a subject, comprising:
(a) preparing a test sample comprising a mucosal fluid from a subject,
(b) bringing the test sample into contact with a target antigen-specific antibody,
(c) adding an agent to suppress inhibition of antigen-antibody binding in the test sample, and
(d) detecting the antigen-antibody binding,
wherein the agent comprises a sulfonic acid compound of the formula R 1 —SO 3 H or a salt thereof, or a quaternary ammonium ion of the formula N + —R 2 R 3 R 4 R 5 or a salt thereof,
wherein R 1 is an unsubstituted straight-chain C 5 -C 20 alkyl group, a straight-chain C 1 -C 20 alkyl group substituted with a phenyl group having a straight-chain C 5 -C 20 alkyl group, or a phenyl group substituted with a straight-chain C 5 -C 20 alkyl group, and
wherein R 2 to R 5 are each independently a straight-chain C 1 -C 20 alkyl group unsubstituted or substituted with a phenyl group, and
wherein the agent is capable of suppressing false positive and/or false negative results of antigen-antibody binding caused by the mucosal fluid in the test sample.

10. The method according to claim 9 ,
wherein the pathogenic disease is a disease caused by a pathogen or a pathogenic microorganism, selected from the group consisting of a foot-and-mouth disease virus, an influenza virus, an adenovirus, a respiratory syncytial virus, a coronavirus, a rabies virus, a Bordetella pertussis , a hemolytic streptococcus , a food poisoning bacterium, a Chlamydia trachomatis , and a mycoplasma.

11. The method according to claim 1 , wherein the agent comprises a straight-chain dodecylbenzenesulfonate, lauryl trimethylammonium chloride, dodecyl trimethylammonium bromide, myristyl trimethylammonium bromide, or benzyl tributylammonium chloride.

12. The method according to claim 9 , wherein the agent comprises a straight-chain dodecylbenzenesulfonate, lauryl trimethylammonium chloride, dodecyl trimethylammonium bromide, myristyl trimethylammonium bromide, or benzyl tributylammonium chloride.

13. The method according to claim 8 , wherein the food poisoning bacterium comprises pathogenic Campylobacter, Salmonella , or Escherichia coli.

14. The method according to claim 10 , wherein the food poisoning bacterium comprises pathogenic Campylobacter, Salmonella , or Escherichia coli.

Assignees

• Nh Foods Ltd
• National Agriculture And Foods Res Organization
• Nat Agriculture & Food Res Org

Inventors

• Takeuchi Eri
• Urayama Kana
• Matsumoto Takashi
• Morioka Kazuki
• Yamakawa Makoto
• Yoshida Kazuo
• Kanno Toru
• Fukai Katsuhiko

Classifications

• G01N33/569 Primary

  for microorganisms, e.g. protozoa, bacteria, viruses · CPC title

• G01N33/56911

  Bacteria · CPC title

• G01N33/56983

  Viruses · CPC title

• G01N1/28

  Preparing specimens for investigation {including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q}(mounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 {; laboratory gas handling apparatus B01L5/00}) · CPC title

• G01N2800/26

  Infectious diseases, e.g. generalised sepsis · CPC title