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DPO multiplex PCR as an alternative to culture and susceptibility testing to detect Helicobacter pylori and its resistance to clarithromycin.

Abstract : 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 Helicobacter 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). CONCLUSION: 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.
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Submitted on : Wednesday, November 2, 2011 - 5:02:26 PM
Last modification on : Friday, October 23, 2020 - 4:33:24 PM
Long-term archiving on: : Thursday, November 15, 2012 - 11:00:38 AM

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Philippe Lehours, Elodie Siffré, Francis Mégraud. DPO multiplex PCR as an alternative to culture and susceptibility testing to detect Helicobacter pylori and its resistance to clarithromycin.. BMC Gastroenterology, BioMed Central, 2011, 11 (1), pp.112. ⟨10.1186/1471-230X-11-112⟩. ⟨inserm-00637709⟩

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