title: Analysis of Helicobacter pylori genotypes in clinical gastric wash samples.
abstract: Helicobacter pylori is a key factor in the development of gastric cancer; indeed, clearance of H. pylori helps prevent gastric cancer. However, the relationship between gastric cancer and the abundance and diversity of H. pylori genotypes in the stomach remains unknown. Here, we present, for the first time, a quantitative analysis of H. pylori genotypes in gastric washes. A method was first developed to assess diversity and abundance by pyrosequencing and analysis of single nucleotide polymorphisms in 23S ribosomal RNA (rRNA), a gene associated with clarithromycin resistance. This method was then validated using arbitrarily mixed plasmids carrying 23S rRNA with single nucleotide polymorphisms. Multiple strains were detected in many of 34 clinical samples, with frequency 24.3 ± 24.2 and 26.3 ± 33.8 % for the A2143G and A2144G strains, respectively. Importantly, results obtained from gastric washes were similar to those obtained from biopsy samples. The method provides opportunities to investigate drug resistance in H. pylori and assess potential biomarkers of gastric cancer risk, and should thus be validated in large-scale clinical trials.
authors.affiliation: Division of Endoscopy, Hokkaido University Hospital, Sapporo, Hokkaido, 060-0808, Japan.|Department of Gastroenterology, Hokkaido University Graduate School of Medicine, Sapporo, Hokkaido, 060-0808, Japan.
authors.forename: Shuichi
authors.initials: S
authors.lastname: Miyamoto
authors.affiliation: Division of Gastroenterology and Hepatology, Department of Internal Medicine, St. Marianna University School of Medicine, Kawasaki, Kanagawa, 216-8511, Japan. ponponta@marianna-u.ac.jp.|Department of Internal Medicine, Kawasaki Rinko General Hospital, 3-13-1 Nakajima Kawasaki-ku, Kawasaki City, Kanagawa, 210-0806, Japan. ponponta@marianna-u.ac.jp.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Watanabe
authors.affiliation: Division of Gastroenterology and Hepatology, Department of Internal Medicine, St. Marianna University School of Medicine, Kawasaki, Kanagawa, 216-8511, Japan.
authors.forename: Ritsuko
authors.initials: R
authors.lastname: Oikawa
authors.affiliation: Division of Endoscopy, Hokkaido University Hospital, Sapporo, Hokkaido, 060-0808, Japan.|Department of Gastroenterology, Hokkaido University Graduate School of Medicine, Sapporo, Hokkaido, 060-0808, Japan.
authors.forename: Shoko
authors.initials: S
authors.lastname: Ono
authors.affiliation: Division of Endoscopy, Hokkaido University Hospital, Sapporo, Hokkaido, 060-0808, Japan.|Department of Gastroenterology, Hokkaido University Graduate School of Medicine, Sapporo, Hokkaido, 060-0808, Japan.|Department of Cancer Preventive Medicine, Graduate School of Medicine, Sapporo, Hokkaido, 060-0808, Japan.
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Mabe
authors.affiliation: Division of Endoscopy, Hokkaido University Hospital, Sapporo, Hokkaido, 060-0808, Japan.
authors.forename: Takahiko
authors.initials: T
authors.lastname: Kudo
authors.affiliation: Division of Gastroenterology and Hepatology, Department of Internal Medicine, St. Marianna University School of Medicine, Kawasaki, Kanagawa, 216-8511, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Yamamoto
authors.affiliation: Division of Gastroenterology and Hepatology, Department of Internal Medicine, St. Marianna University School of Medicine, Kawasaki, Kanagawa, 216-8511, Japan.
authors.forename: Fumio
authors.initials: F
authors.lastname: Itoh
authors.affiliation: Division of Endoscopy, Hokkaido University Hospital, Sapporo, Hokkaido, 060-0808, Japan.
authors.forename: Mototsugu
authors.initials: M
authors.lastname: Kato
authors.affiliation: Department of Gastroenterology, Hokkaido University Graduate School of Medicine, Sapporo, Hokkaido, 060-0808, Japan.
authors.forename: Naoya
authors.initials: N
authors.lastname: Sakamoto
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D001706:Biopsy; D017291:Clarithromycin / Q000494:pharmacology; D004269:DNA, Bacterial / Q000235:genetics; D004352:Drug Resistance, Microbial / Q000235:genetics; D005260:Female; D005756:Gastritis / Q000382:microbiology; D005773:Gastroscopy; D005838:Genotype; D016481:Helicobacter Infections / Q000382:microbiology; D016480:Helicobacter pylori / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008297:Male; D008875:Middle Aged; D020641:Polymorphism, Single Nucleotide; D012329:RNA, Bacterial / Q000235:genetics; D012338:RNA, Ribosomal, 23S; D017423:Sequence Analysis, RNA; D013270:Stomach / Q000382:microbiology*; D007507:Therapeutic Irrigation
chemical_list: D004269:DNA, Bacterial; D012329:RNA, Bacterial; D012338:RNA, Ribosomal, 23S; D017291:Clarithromycin
keywords: 23S rRNA; Gastric wash; Helicobacter pylori; Pyrosequencing
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