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JP-7854593-B2 - Method for detecting Helicobacter Swiss antibodies using whole bacterial cells

JP7854593B2JP 7854593 B2JP7854593 B2JP 7854593B2JP-7854593-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 antibodies that bind to H. swiss whole cells in a subject-derived sample, including at least H. swiss whole cells and anti-Ig antibodies.
  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 antibodies that bind to anti-H. Swiss whole cells in a sample derived from a subject, comprising the following steps: (a) a step of contacting a sample derived from a subject with whole H. swiss bacteria, and (b) a step of detecting antibodies in the sample that have been bound to whole H. swiss bacteria.
  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 whole H. suisse cells. 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 as the sample and H. swiss whole cells and HsvA antigen peptide as the antigens.This graph shows the results of ELISA using human serum as the sample and H. swiss whole cells and HsvA antigen peptide as the antigens.This figure shows the measured values of ELISA results using human serum (3600-fold dilution) as the sample and whole H. pylori or whole H. swiss bacteria as the antigen.This figure shows the results of ELISA using human serum (3600-fold dilution) as the sample and either whole H. pylori cells (Figure 4A) or whole H. swiss bacteria (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 whole H. swiss cells 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 either whole H. swiss cells 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 all H. suissian bacteria in a biological sample of a subject, and a reagent for detecting said antibodies. The present invention also relates to a method for determining the presence of H. suissian in a subject, or a method for detecting H. suissian infection in a subject. Alternatively, it relates to a method for obtaining auxiliary data for diagnosing H. suissian infection in a subject. (H. Switzerland: Helicobacter suiis) In this specification, "H. suisse" refers to the H. suisse species included in NHPH (Non-Helicobacter pylori helicobacters; Helicobacter other than H. pylori) or Helicobacter heilmannii sensu lato (Helicobacter heilmannii in the broad sense) among the more than 50 reported species of Helicobacter bacteria (Non-Patent Literature 3). H. su