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Journal of Clinical Microbiology, August 2003, p. 3636-3640, Vol. 41, No. 8
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.8.3636-3640.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Detection of Clarithromycin-Resistant Helicobacter pylori in Stool Samples

Carla Fontana,1,2* Marco Favaro,3 Antonio Pietroiusti,4 Enrico Salvatore Pistoia,1 Alberto Galante,4,5 and Cartesio Favalli1,2

Department of Experimental Medicine and Biochemical Sciences,1 Department of Biology,3 Department of Internal Medicine, "Tor Vergata" University of Rome,4 Clinical Microbiology Laboratories, Policlinic of Tor Vergata, 00133 Rome,2 Clinica San Raffaele, Velletri, Italy5

Received 29 January 2003/ Returned for modification 30 March 2003/ Accepted 24 May 2003

The recognition of the role of Helicobacter pylori in gastric diseases has led to the widespread use of antibiotics in the eradication of this pathogen. The most advocated therapy, triple therapy, often includes clarithromycin. It is well known that clarithromycin resistance is one of the major causes of eradication failure. The development of a rapid noninvasive technique that could easily be performed on fecal samples and that could also provide information about the antibiotic resistance of this microorganism is therefore advisable. Previous findings have demonstrated that clarithromycin resistance is due to a single point mutation in the 23S rRNA. All the mutations described have been associated with specific restriction sites, namely BsaI (A2143G), MboII (A2142C/G), and HhaI (T2717C). On this basis we have developed a new method, a seminested PCR, allowing screening for clarithromycin resistance of H. pylori directly on stool samples. This method furnished a 783-bp fragment of the 23S rRNA, which was subsequently digested by MboII, BsaI, and HhaI, in order to identify single point mutations associated with clarithromycin resistance. Of a total of 283 stool samples examined, 125 were H. pylori positive and two of them were shown to contain clarithromycin-resistant strains due to the presence of a mutation at position 2717, whereas no PCR products contained mutations at position 2142 or 2143. In order to evaluate the reliability of the new system, we compared the results of restriction analysis of the PCR products with the MICs shown by the H. pylori isolates by culturing gastric biopsies from the same patients.

* Corresponding author. Mailing address: Dept. of Experimental Medicine and Biochemical Sciences, "Tor Vergata" University of Rome, Via Montepellier 1, 00133 Rome, Italy. Phone: 039 6 20902158. Fax: 039 6 20902078. E-mail: fontana{at}med.uniroma2.it.

Journal of Clinical Microbiology, August 2003, p. 3636-3640, Vol. 41, No. 8
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.8.3636-3640.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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  • Moder, K.-A., Layer, F., Konig, W., Konig, B. (2007). Rapid screening of clarithromycin resistance in Helicobacter pylori by pyrosequencing. J Med Microbiol 56: 1370-1376 [Abstract] [Full Text]  
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