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Journal of Clinical Microbiology, November 2009, p. 3600-3607, Vol. 47, No. 11
0095-1137/09/$08.00+0     doi:10.1128/JCM.00744-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Evaluation of a New Test, GenoType HelicoDR, for Molecular Detection of Antibiotic Resistance in Helicobacter pylori{triangledown}

Emmanuelle Cambau,1,2* Vera Allerheiligen,3 Céline Coulon,1 Céline Corbel,2 Christine Lascols,1 Lionel Deforges,1,2 Claude-James Soussy,1,2 Jean-Charles Delchier,1,4 and Francis Megraud5,6

Université Paris 12, Créteil, F-94000, France,1 AP-HP, Hôpital Henri-Mondor, Laboratoire de Bactériologie-Virologie-Hygiène, Créteil, F-94000, France,2 Hain Lifesciences, Nehren, Germany,3 AP-HP, Hôpital Henri-Mondor, Service d'Hépato-Gastroentérologie, Créteil, F-94000, France,4 Centre National de Référence des Campylobacter et Helicobacter, Université Victor Segalen Bordeaux 2, Bordeaux, France,5 INSERM U853, 33076 Bordeaux Cedex, France6

Received 13 April 2009/ Returned for modification 21 June 2009/ Accepted 8 September 2009

The eradication rate of Helicobacter pylori by standard therapy is decreasing due to antibiotic resistance, mainly to clarithromycin. Our aim was to provide a new molecular test to guide the treatment of new and relapsed cases. We first studied 126 H. pylori strains for phenotypic (MIC) and genotypic resistance to clarithromycin (rrl mutation) and levofloxacin (gyrA mutation) and then developed a DNA strip genotyping test on the basis of the correlation results and literature data. Clinical strains (n = 92) and gastric biopsy specimens containing H. pylori (n = 105) were tested blindly with the new molecular test GenoType HelicoDR. The presence of mutations or the absence of hybridization with wild-type sequences was predictive, in rrl for clarithromycin resistance in 91 cases (mostly the A2147G mutation) and in gyrA for levofloxacin resistance in 58 cases (mutations at codon 87 or 91). Genotyping revealed a mix of genotypes in 33% of the cases, reflecting a coinfection or selection for resistant mutants. The sensitivity and specificity of detecting resistance were 94% and 99% for clarithromycin and 87% and 98.5% for levofloxacin, respectively. The concordance scores were 0.96 for clarithromycin and 0.94 for levofloxacin. With global resistance rates of 46% for clarithromycin and 25% for levofloxacin, which were observed for consecutive positive biopsy specimens from 2007 and 2008, the positive and negative predictive values for detecting resistance were 99% and 94% for clarithromycin and 96% and 96% for fluoroquinolone. GenoType HelicoDR is efficient at detecting mutations predictive of antibiotic resistance in H. pylori when applied to strains or directly to gastric biopsy specimens.

* Corresponding author. Present address: Laboratoire de Bactériologie-Virologie-Hygiène, Hôpital Saint Louis, 1 Avenue Claude Vellefaux, 75018 Paris Cedex, France. Phone: 33 1 42499348. Fax: 33 1 42499200. E-mail: emmanuelle.cambau{at}sls.aphp.fr

{triangledown} Published ahead of print on 16 September 2009.

Journal of Clinical Microbiology, November 2009, p. 3600-3607, Vol. 47, No. 11
0095-1137/09/$08.00+0     doi:10.1128/JCM.00744-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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