title: DPO multiplex PCR as an alternative to culture and susceptibility testing to detect Helicobacter pylori and its resistance to clarithromycin.
abstract: BACKGROUND
  Macrolide resistance in Helicobacter pylori is the major risk factor for treatment failure when using a proton pump inhibitor-clarithromycin containing therapy. Macrolide resistance is due to a few mutations on the 23S ribomosal subunit encoded by the 23S rRNA gene. The present study aimed at investigating the performance of the dual priming oligonucleotide (DPO)-PCR kit named Seeplex® ClaR-H. pylori ACE detection designed to detect H. pylori and two types of point mutations causing clarithromycin resistance in H. pylori.
  
  
  METHODS
  The performance of Seeplex® ClaR-H. pylori ACE detection was evaluated on 127 gastric biopsies in comparison to conventional bacterial culture followed by the determination of susceptibility to clarithromycin by E-test, as well as by an in-house real-time PCR using a fluorescence resonance energy transfer (FRET) technology.
  
  
  RESULTS
  Considering culture as the reference test, the sensitivity of DPO-PCR and real-time FRET-PCR was 97.7% and 100% while specificity was 83.1% and 80.7%, respectively. However, both PCR were concordant in detecting 14 H. pylori positive cases which were negative by culture. Globally, E-test and DPO-PCR were concordant with regard to clarithromycin susceptibility in 95.3% of the cases (41/43), while real-time FRET-PCR and DPO-PCR were concordant in 95% (57/60).
  
  
  CONCLUSIONS
  The DPO-PCR is an interesting tool to detect H. pylori on gastric biopsies and to study its susceptibility to clarithromycin in laboratories that cannot perform real-time PCR assays.
authors: Université de Bordeaux, Centre National de Référence des Campylobacters et des Hélicobacters, 146 rue Léo Saignat, 33000 Bordeaux, France.
authors: Philippe
authors: P
authors: Lehours
authors: Elodie
authors: E
authors: Siffré
authors: Francis
authors: F
authors: Mégraud
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001706:Biopsy; D017291:Clarithromycin / Q000494:pharmacology; D017931:DNA Primers / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D016481:Helicobacter Infections / Q000188:drug therapy* / Q000382:microbiology; D016480:Helicobacter pylori / Q000235:genetics*; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009841:Oligonucleotides / Q000235:genetics; D017354:Point Mutation; D060888:Real-Time Polymerase Chain Reaction; D013270:Stomach / Q000473:pathology
chemical_list: D000900:Anti-Bacterial Agents; D017931:DNA Primers; D009841:Oligonucleotides; D017291:Clarithromycin