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Antimicrobial Agents and Chemotherapy, September 2002, p. 2996-3000, Vol. 46, No. 9
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.9.2996-3000.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

16S rRNA Mutation-Mediated Tetracycline Resistance in Helicobacter pylori

Monique M. Gerrits,1 Marcel R. de Zoete,1 Niek L. A. Arents,2 Ernst J. Kuipers,1 and Johannes G. Kusters1*

Department of Gastroenterology and Hepatology, Erasmus MC—University Medical Center Rotterdam, Rotterdam,1 Medical Microbiology, Regional Public Health Laboratory Groningen/Drenthe, Groningen, The Netherlands2

Received 12 February 2002/ Returned for modification 18 April 2002/ Accepted 29 May 2002

Most Helicobacter pylori strains are susceptible to tetracycline, an antibiotic commonly used for the eradication of H. pylori. However, an increase in incidence of tetracycline resistance in H. pylori has recently been reported. Here the mechanism of tetracycline resistance of the first Dutch tetracycline-resistant (Tetr) H. pylori isolate (strain 181) is investigated. Twelve genes were selected from the genome sequences of H. pylori strains 26695 and J99 as potential candidate genes, based on their homology with tetracycline resistance genes in other bacteria. With the exception of the two 16S rRNA genes, none of the other putative tetracycline resistance genes was able to transfer tetracycline resistance. Genetic transformation of the Tets strain 26695 with smaller overlapping PCR fragments of the 16S rRNA genes of strain 181, revealed that a 361-bp fragment that spanned nucleotides 711 to 1071 was sufficient to transfer resistance. Sequence analysis of the 16S rRNA genes of the Tetr strain 181, the Tets strain 26695, and four Tetr 26695 transformants showed that a single triple-base-pair substitution, AGA926-928->TTC, was present within this 361-bp fragment. This triple-base-pair substitution, present in both copies of the 16S rRNA gene of all our Tetr H. pylori transformants, resulted in an increased MIC of tetracycline that was identical to that for the Tetr strain 181.

* Corresponding author. Mailing address: Department of Gastroenterology and Hepatology, Rm. L457, Erasmus MC - University Medical Center Rotterdam, Dr. Molewaterplein 40, 3015 GD Rotterdam, The Netherlands. Phone: 31-10-4632982. Fax: 31-10-4632793. E-mail: Kusters{at}mdl.azr.nl.

Antimicrobial Agents and Chemotherapy, September 2002, p. 2996-3000, Vol. 46, No. 9
0066-4804/02/$04.00+0     DOI: 10.1128/AAC.46.9.2996-3000.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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