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JP-7854592-B2 - A method for detecting Helicobacter Swiss antibodies using the extracellular membrane fraction.

JP7854592B2JP 7854592 B2JP7854592 B2JP 7854592B2JP-7854592-B2

Inventors

  • 松井 英則
  • 林原 絵美子
  • 鈴木 仁人
  • 高橋 崇道
  • 石井 明

Assignees

  • 学校法人北里研究所
  • 国立健康危機管理研究機構
  • デンカ株式会社

Dates

Publication Date
20260507
Application Date
20230125
Priority Date
20220125

Claims (11)

  1. A reagent for detecting the H. Swiss outer membrane fraction of a subject-derived sample, or antibodies that bind to proteins contained in the outer membrane fraction, including at least the H. Swiss outer membrane fraction, or proteins and anti-Ig antibodies contained in the outer membrane fraction.
  2. The reagent according to claim 1, characterized in that the test subject is a mammal.
  3. The reagent according to claim 1, characterized in that the sample is derived from blood.
  4. Furthermore, the reagent according to claim 1 is characterized by also detecting antibodies that bind to H. pylori or antigens of H. pylori bacterial components in a subject-derived sample, including H. pylori bacterial components or antibodies against H. pylori bacterial components.
  5. The reagent according to claim 1, which is an ELISA or immunochromatographic reagent.
  6. A method characterized by detecting an antibody that binds to the anti-H. Swiss outer membrane fraction or a protein contained in the outer membrane fraction of a subject-derived sample, comprising the following steps: (a) a step of contacting a sample derived from a subject with the H. suisse outer membrane fraction or proteins contained in the outer membrane fraction, and (b) a step of detecting antibodies in the sample that are bound to the H. suisse outer membrane fraction or proteins contained in the outer membrane fraction.
  7. The method according to claim 6, characterized in that the subject is a mammal.
  8. The method according to claim 6, characterized in that the sample is derived from blood.
  9. Furthermore, the method according to claim 6 is characterized by detecting antibodies that bind to H. pylori in a sample derived from the subject, comprising the following steps: (a) a step of contacting a sample derived from a subject with H. pylori cell components, and (b) a step of detecting antibodies in the sample that are bound to H. pylori cell components.
  10. Furthermore, the method according to claim 6 is characterized by also detecting antigens of H. pylori cell components in a sample derived from the subject, including the following steps: (a) a step of contacting a sample derived from a subject with an antibody against H. pylori cell components, and (b) a step of detecting the antigen of the H. pylori cell component in the sample that has been bound to the antibody against H. pylori cell components.
  11. A method for obtaining supplementary data for detecting H. swiss infection in a subject , comprising detecting antibodies by measurement using a reagent described in any one of claims 1 to 5 and/or by the method described in any one of claims 6 to 10.

Description

This invention relates to a method for measuring antibodies against Helicobacter suisse using the outer membrane fraction of H. suisse. Currently, infection with Helicobacter pylori (a microaerophilic, Gram-negative spiral bacterium that parasitizes the human stomach, hereinafter referred to as "H. pylori") is known to be involved in chronic gastritis, gastric ulcers and duodenal ulcers, gastric cancer, gastric mucosa-associated lymphoid tissue (MALT) lymphoma, and diffuse large B-cell lymphoma. Common methods for testing H. pylori infection include isolation culture, urea breath test (UBT), measurement of H. pylori antibody titers in serum and urine (ELISA and latex agglutination), measurement of H. pylori antigen in stool (immunochromatography), and rapid urease test (RUT) of gastric biopsy samples. However, in recent years, with the decline in H. pylori infection rates due to the widespread diagnosis and eradication of H. pylori, NHPH (Non-Helicobacter pylori helicobacters; Helicobacters other than H. pylori) has become a problem as a Helicobacter species that induces serious gastric diseases in humans. NHPH includes Helicobacter swissii (hereinafter, H. swissii), H. bizzozeronnii, H. felis, H. salmonis, H. ailurogasticus, H. cynogasticus, H. baculiformis, H. mustelae, H. acinonychls, H. cetorum, and H. This includes *H. heilmannii*, of which the majority found in human stomachs are *H. swisii*. H. pylori only infects primates, and infection is thought to occur only through contact with close relatives during infancy, whereas H. swiss can be transmitted from animals such as pigs and monkeys, regardless of age. H. suiss parasitizes the stomachs of monkeys approximately 100,000 years ago. Later, 15,000 years ago, it began infecting pigs. With the domestication of pigs, the infection spread explosively, eventually infecting humans (Non-Patent Literature 1). MLST (Multi-Locus Sequencing Typing) analysis of 181 H. suiss gene strains isolated from around the world revealed that independent clusters formed depending on the infected host animal. However, strains isolated from humans did not form independent clusters and were included in the pig cluster, leading to the conclusion that pigs were the source of human infection (Non-Patent Literature 2). Therefore, infection diagnosis and disinfection agents are needed for both pigs and humans. Regarding H. swiss infection testing methods, measurement of H. swiss antibody titers in serum (ELISA and latex agglutination method) has been reported (Patent Documents 1 and 2). Japanese Patent Publication No. 2016-10331International Publication WO2019/225639 Flahou et al. , ISM J. , Jan; 12(1):77-86. doi:10.1038/ismej. 2017.145E. Rimbara et al. , Proc Natl Acad Sci USA 2021 Vol. 118 Issue 13 e2026337118. doi:10.1073/pnas. 2026337118.Haesebrouck F. et al. , Helicobacter, 16(4), 339-340, 2011, doi:10.1111/j. 1523-5378.2011.00849. xA. D. Augustin, et al. , Front. Med. , 2019, 6, 188, doi:10.3389/fmed. 2019.00188 (https://www.ncbi.nlm.nih.gov/pubmed/31555648) This figure shows the measured values of ELISA results using human serum (3600-fold dilution) as the sample and either whole H. swiss cells or the outer membrane fraction of H. swiss as the antigen.This figure shows the results of ELISA using human serum (3600-fold dilution) as the sample and either whole H. swiss cells (Figure 2A) or the outer membrane fraction of H. swiss (Figure 2B) as the antigen.This figure shows the measured values of ELISA results using mouse serum (3600-fold dilution) as the sample and either whole H. swiss cells or the outer membrane fraction of H. swiss as the antigen.This figure shows the results of ELISA using mouse serum (3600-fold dilution) as the sample and either whole H. swiss cells (Figure 4A) or the outer membrane fraction of H. swiss (Figure 4B) as the antigen.This figure shows the measured values of ELISA results using human serum (3600-fold dilution) as the sample and either the H. pylori outer membrane fraction or the H. swiss outer membrane fraction as the antigen.This figure shows the results of ELISA using human serum (3600-fold dilution) as the sample and either the H. pylori outer membrane fraction (Figure 6A) or the H. swiss outer membrane fraction (Figure 6B) as the antigen.This figure shows the measured values of ELISA results using human serum (3600-fold dilution) as the sample and H. swiss outer membrane fraction or partial peptides of H. swiss HsVA (SEQ ID NOs: 1-5) as the antigen.This figure shows the measured values of ELISA results using human serum (3600-fold dilution) as the sample and H. swiss outer membrane fraction or partial peptides of H. swiss HsVA (SEQ ID NOs. 6-11) as the antigen. The method of the present invention will be described below. The present invention relates to a method for detecting antibodies against the outer membrane fraction of H. suisse bacteria, or proteins contained in the outer membrane fraction, in a biological sample of a subject, and a