Dual-priming oligonucleotide-based multiplex PCR for the detection of Helicobacter pylori and determination of clarithromycin resistance with gastric biopsy specimens

Helicobacter. 2009 Feb;14(1):22-8. doi: 10.1111/j.1523-5378.2009.00654.x.


Background: Assessment of Helicobacter pylori (H. pylori) clarithromycin resistance has rarely been performed routinely despite an increasing resistance rate. Our aim was to develop and evaluate the use of dual-priming oligonucleotide (DPO)-based multiplex polymerase chain reaction (PCR) to detect point mutations in the 23S rRNA gene responsible for clarithromycin resistance of H. pylori.

Materials and methods: Gastric biopsy specimens from 212 untreated patients with dyspepsia were examined by culture, histology, and DPO-based multiplex PCR. A disk diffusion test and E-test were used for performing phenotypic antibiotic susceptibility tests.

Results: Among the biopsy specimens tested, 22.2% (47/212), 42.5% (90/212), and 41.5% (88/212) of the specimens were classified as H. pylori positive by culture, histology, and DPO-based multiplex PCR, respectively. Among 96 strains identified by either culture or DPO-based multiplex PCR, 80 strains were clarithromycin-susceptible and 16 strains (16.7%) were clarithromycin-resistant. There was 94.1% (32/34) concordance between phenotypic susceptibility tests and DPO-based multiplex PCR. In two patients with discrepant results, only DPO-based multiplex PCR detected clarithromycin-resistant strains. DPO-based multiplex PCR identified additional 49 clarithromycin-resistant or clarithromycin-susceptible H. pylori among 165 culture-negative specimens.

Conclusions: DPO-based multiplex PCR can be used as a practical method for the detection of H. pylori infection and the determination of clarithromycin susceptibility in addition to phenotypic antimicrobial susceptibility tests.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Anti-Bacterial Agents / pharmacology*
  • Biopsy
  • Clarithromycin / pharmacology*
  • DNA Primers / genetics
  • DNA, Bacterial / genetics
  • DNA, Ribosomal / genetics
  • Drug Resistance, Bacterial*
  • Female
  • Helicobacter Infections / microbiology*
  • Helicobacter Infections / pathology
  • Helicobacter pylori / drug effects
  • Helicobacter pylori / genetics
  • Helicobacter pylori / isolation & purification*
  • Humans
  • Korea
  • Male
  • Middle Aged
  • Mutation
  • Polymerase Chain Reaction / methods*
  • RNA, Ribosomal, 23S / genetics


  • Anti-Bacterial Agents
  • DNA Primers
  • DNA, Bacterial
  • DNA, Ribosomal
  • RNA, Ribosomal, 23S
  • Clarithromycin