Tumor Biology

, Volume 37, Issue 8, pp 10123–10132 | Cite as

Analysis of Helicobacter pylori genotypes in clinical gastric wash samples

  • Shuichi Miyamoto
  • Yoshiyuki Watanabe
  • Ritsuko Oikawa
  • Shoko Ono
  • Katsuhiro Mabe
  • Takahiko Kudo
  • Hiroyuki Yamamoto
  • Fumio Itoh
  • Mototsugu Kato
  • Naoya Sakamoto
Original Article


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.


Helicobacter pylori Gastric wash 23S rRNA Pyrosequencing 



This work was supported by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan to Y.W. (#25460937).

Authors’ contributions

S.M., Y.W., and H.Y. conceived the study, designed and executed experiments, analyzed data, prepared figures and tables, and wrote the manuscript. S.O. analyzed data, and K.M. helped write the manuscript. R.O. and S.M. executed experiments and analyzed data. S.O. and K.M. obtained clinical samples. M.K. designed experiments, analyzed data, and helped write the manuscript. F.I. provided intellectual support. T.K., M.K., and N.S. supervised all aspects of the study.

Compliance with ethical standards

The study was conducted in accordance with all rules and regulations of the Hokkaido University Institutional Review Board (#014-0459), and informed consent was obtained from each patient.

Conflicts of interest



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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Shuichi Miyamoto
    • 1
    • 2
  • Yoshiyuki Watanabe
    • 4
    • 5
  • Ritsuko Oikawa
    • 4
  • Shoko Ono
    • 1
    • 2
  • Katsuhiro Mabe
    • 1
    • 2
    • 3
  • Takahiko Kudo
    • 1
  • Hiroyuki Yamamoto
    • 4
  • Fumio Itoh
    • 4
  • Mototsugu Kato
    • 1
  • Naoya Sakamoto
    • 2
  1. 1.Division of EndoscopyHokkaido University HospitalSapporoJapan
  2. 2.Department of GastroenterologyHokkaido University Graduate School of MedicineSapporoJapan
  3. 3.Department of Cancer Preventive MedicineGraduate School of MedicineSapporoJapan
  4. 4.Division of Gastroenterology and Hepatology, Department of Internal MedicineSt. Marianna University School of MedicineKawasakiJapan
  5. 5.Department of Internal MedicineKawasaki Rinko General HospitalKawasaki CityJapan

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