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Journal of Clinical Microbiology, February 2007, p. 303-305, Vol. 45, No. 2
0095-1137/07/$08.00+0     doi:10.1128/JCM.01997-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Rapid Detection of Point Mutations Conferring Resistance to Fluoroquinolone in gyrA of Helicobacter pylori by Allele-Specific PCR{triangledown}

Toshihiro Nishizawa,1 Hidekazu Suzuki,1* Ayako Umezawa,2 Hiroe Muraoka,3 Eisuke Iwasaki,1 Tatsuhiro Masaoka,4 Intetsu Kobayashi,3 and Toshifumi Hibi1

Department of Internal Medicine,1 Center for Integrated Medical Research, Keio University School of Medicine,2 Mitsubishi Kagaku Bio-Clinical Laboratories,3 Department of Gastroenterology, Eiju Hospital, Tokyo, Japan4

Received 27 September 2006/ Returned for modification 18 October 2006/ Accepted 8 November 2006

Helicobacter pylori strains with reduced susceptibility to fluoroquinolones have a mutation at either codon 87 Asn or 91 Asp of the gyrA gene. A rapid test based on an allele-specific PCR (AS-PCR) was designed to detect the gyrA mutations. Clinical H. pylori isolates were obtained from the stomachs of 51 patients with H. pylori infections who showed treatment failure. The MICs of gatifloxacin (GAT) were determined by the agar dilution method. Identical genotyping results were obtained with AS-PCR and conventional PCR. The gyrA mutations of H. pylori causing reduced susceptibility to fluoroquinolones could be detected successfully by this method. A significant association was observed between the presence of mutations, as detected by AS-PCR, and the resistance of the strains to GAT. Moreover, genotyping by AS-PCR took less than 3 to 4 h. The AS-PCR method for the detection of gyrA mutations in H. pylori is useful for easy identification of fluoroquinolone-resistant strains of H. pylori.

* Corresponding author. Mailing address: Department of Internal Medicine, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan. Phone: 81-3-5363-3914. Fax: 81-3-5363-3967. E-mail: hsuzuki{at}sc.itc.keio.ac.jp.

{triangledown} Published ahead of print on 22 November 2006.

Journal of Clinical Microbiology, February 2007, p. 303-305, Vol. 45, No. 2
0095-1137/07/$08.00+0     doi:10.1128/JCM.01997-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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