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Journal of Antimicrobial Chemotherapy (2002) 49, 549-552
© 2002 The British Society for Antimicrobial Chemotherapy

Brief report

Use of a rapid mismatch PCR method to detect gyrA and parC mutations in ciprofloxacin-resistant clinical isolates of Escherichia coli

Yan Zhi Qianga, Tong Qina, Wang Fub, Wu Pei Chengb, Yang Sheng Lia and Gong Yia,*

a Shanghai Research Center of Biotechnology, SIBS, Chinese Academy of Sciences, 500 Cao Bao Road, Shanghai 200233; b Institute of Antibiotics, Medical Center of Fu Dan University, Shanghai, China

Four amino acid substitutions, two in GyrA and two in ParC subunits of DNA gyrase and topoisomerase IV, respectively, are commonly responsible for fluoroquinolone resistance in Escherichia coli. In this study, an economical and time-efficient mismatch amplification mutation assay (MAMA) PCR was developed to detect mutations in the chromosomal gyrA and parC genes causing these substitutions. One hundred and twenty-one clinical E. coli isolates were tested by this assay, and the results confirmed that accumulation of amino acid alterations in GyrA and ParC correlates closely with stepwise increases in the MIC of ciprofloxacin.

* Corresponding author. Tel: +86-21-6436-9607; Fax: +86-21-6470-0244; E-mail: yigong{at}srcb.ac.cn


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