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

BMC Gastroenterology

Research Article

Elodie Siffré¹  Francis Mégraud²  Philippe Lehours² 

[1] Centre National de Référence des Campylobacters et des Hélicobacters, Université de Bordeaux, 146 rue Léo Saignat, 33000, Bordeaux, France;Centre National de Référence des Campylobacters et des Hélicobacters, Université de Bordeaux, 146 rue Léo Saignat, 33000, Bordeaux, France;CHU de Bordeaux, Hôpital Pellegrin, Laboratoire de Bactériologie, Place Amélie Raba Léon, 33076, Bordeaux cedex, France;INSERM U853, 33000, Bordeaux, France;

关键词: Clarithromycin;    Fluorescence Resonance Energy Transfer;    Gastric Biopsy;    Macrolide Resistance;    Clarithromycin Resistance;   

DOI  :  10.1186/1471-230X-11-112

 received in 2011-07-26, accepted in 2011-10-17,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundMacrolide 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.MethodsThe 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.ResultsConsidering 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).ConclusionThe 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.

【 授权许可】

Unknown   
© Lehours et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.