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Antimicrobial Agents and Chemotherapy, December 2002, p. 3765-3769, Vol. 46, No. 12
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.12.3765-3769.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

New Site of Modification of 23S rRNA Associated with Clarithromycin Resistance of Helicobacter pylori Clinical Isolates

Carla Fontana,1,2* Marco Favaro,3 Silvia Minelli,1,2 Anna Angela Criscuolo,1,2 Antonio Pietroiusti,4 Alberto Galante,4 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, Italy2

Received 1 April 2002/ Returned for modification 10 July 2002/ Accepted 23 August 2002

Resistance of Helicobacter pylori to clarithromycin occurs with a prevalence ranging from 0 to 15%. This has an important clinical impact on dual and triple therapies, in which clarithromycin seems to be the better choice to achieve H. pylori eradication. In order to evaluate the possibility of new mechanisms of clarithromycin resistance, a PCR assay that amplified a portion of 23S rRNA from H. pylori isolates was used. Gastric tissue biopsy specimens from 230 consecutive patients were cultured for H. pylori isolation. Eighty-six gastric biopsy specimens yielded H. pylori-positive results, and among these 12 isolates were clarithromycin resistant. The latter were studied to detect mutations in the 23S rRNA gene. Sequence analysis of the 1,143-bp PCR product (portion of the 23S rRNA gene) did not reveal mutation such as that described at position 2142 to 2143. On the contrary, our findings show, for seven isolates, a T-to-C transition at position 2717. This mutation conferred a low level of resistance, equivalent to the MIC for the isolates, selected using the E-test as well as using the agar dilution method: 1 µg/ml. Moreover, T2717C transition is located in a highly conserved region of the 23S RNA associated with functional sites: domain VI. This fact has a strong effect on the secondary structure of the 23S RNA and on its interaction with macrolide. Mutation at position 2717 also generated an HhaI restriction site; therefore, restriction analysis of the PCR product also permits a rapid detection of resistant isolates.

* Corresponding author. Mailing address: Department of Experimental Medicine and Biochemical Sciences, "Tor Vergata" University of Rome, Via Montepellier 1, 00133 Rome, Italy. Phone: 039-6-20902158. Fax: 039-6-20900676. E-mail: fontanac{at}med.uniroma2.it.

Antimicrobial Agents and Chemotherapy, December 2002, p. 3765-3769, Vol. 46, No. 12
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.12.3765-3769.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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