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Detection of four clarithromycin resistance point mutations in Helicobacter pylori: comparison of real-time PCR and PCR-RFLP methods

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Abstract

Helicobacter pylori infection can be eradicated in up to 90 % of patients using current combination of triple therapies, of which clarithromycin is a key component. The development of clarithromycin resistance in H. pylori is recognized as a significant contributing factor in treatment failure. The aim of this study is to compare two fast and direct diagnostic methods to evaluate clarithromycin resistance in gastric biopsy specimens. This cross-sectional descriptive study was performed on 150 gastric biopsy specimens. Analysis for clarithromycin resistance was performed by real-time polymerase chain reaction (PCR) assay for A2144G, A2143G, A2143C, and A2142G point mutations by specific probes. Also, A2142G and A2143G point mutations were detected by PCR-RFLP method and using BsaI and MboII restriction enzymes. Out of 150 samples, 96 (64 %) clarithromycin-sensitive- and 54 (36 %) clarithromycin-resistant strains were detected by TaqMan real-time PCR assay. Thirty-seven (24.67 %) resistant strains were detected by PCR-RFLP method. Results showed that real-time PCR assay has enough accuracy to identify clarithromycin resistance and their mutations in a short time.

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Acknowledgments

The authors offer their thanks and appreciation to the Islamic Azad University, Jahrom and Shahrekord branches for their executive protection of this project.

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Authors and Affiliations

  1. Department of Microbiology, Jahrom Branch, Islamic Azad University, Jahrom, Iran

    Mohammad Kargar

  2. Biotechnology Research Center, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran

    Abbas Doosti

  3. Department of Microbiology, Jahrom Branch, Young Researcher’s Club, Islamic Azad University, Jahrom, Iran

    Sadegh Ghorbani-Dalini

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  1. Mohammad Kargar
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  2. Abbas Doosti
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  3. Sadegh Ghorbani-Dalini
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Correspondence to Mohammad Kargar.

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Kargar, M., Doosti, A. & Ghorbani-Dalini, S. Detection of four clarithromycin resistance point mutations in Helicobacter pylori: comparison of real-time PCR and PCR-RFLP methods. Comp Clin Pathol 22, 1007–1013 (2013). https://doi.org/10.1007/s00580-012-1519-1

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  • DOI: https://doi.org/10.1007/s00580-012-1519-1

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