This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ozeki, S. Articles by Kawada, Y. Search for Related Content PubMed PubMed Citation Articles by Ozeki, S. Articles by Kawada, Y.

 Previous Article  |  Next Article 

Journal of Clinical Microbiology, Sep 1997, 2315-2319, Vol 35, No. 9
Copyright © 1997 by the American Society for Microbiology. All rights reserved.

Development of a rapid assay for detecting gyrA mutations in Escherichia coli and determination of incidence of gyrA mutations in clinical strains isolated from patients with complicated urinary tract infections

S Ozeki, T Deguchi, M Yasuda, M Nakano, T Kawamura, Y Nishino and Y Kawada
Department of Urology, Gifu University School of Medicine, Gifu City, Japan.

The MICs of ofloxacin for 743 strains of Escherichia coli isolated from 1988 to 1994 were determined by testing. The strains were from patients with urinary tract infections complicated by functional or anatomical disorders of the urinary tract. Those determined to be ofloxacin resistant (MIC, > or =12.5 microg/ml) comprised 3 of 395 strains (1.3%) from the 1988 to 1990 group, 2 of 166 strains (1.2%) from the 1991 to 1992 group, and 7 of 182 strains (3.8%) from the 1993 to 1994 group. The incidence of resistant strains increased significantly during this period. The percentage of isolates with moderately decreased susceptibilities to ofloxacin (MIC, 0.39 to 3.13 microg/ml) also rose during the same period. To determine the incidence of gyrA mutations in urinary-tract-derived strains of E. coli, we developed a simple and rapid assay based on PCR amplification of the region of the gyrA gene containing the mutation sites followed by digestion of the PCR product with a restriction enzyme. Using this assay, we examined all 182 strains isolated in 1993 and 1994 for the presence of mutations at Ser- 83 and Asp-87 in the gyrA gene. Of these strains, 33 (18.1%) had mutations in the gyrA gene. The incidences of mutations at Ser-83, at Asp-87, and at both codons were 10.4 (19 strains), 4.4 (8 strains), and 3.3% (6 strains), respectively. To determine the correlation of the mutations in the gyrA gene with susceptibilities to quinolones (nalidixic acid, ofloxacin, norfloxacin, and ciprofloxacin), we further examined 116 strains for which the MICs of ofloxacin were > or =0.2 microg/ml that were chosen from the isolates in the 1988 to 1992 group. The MICs of nalidixic acid for the strains without mutations at either Ser-83 or Asp-87 were < or =25 microg/ml, whereas those for the strains with single mutations or double mutations were from 50 to >800 microg/ml. For the fluoroquinolones, significant differences in the distributions of the MICs were observed among the strains without mutations, with single mutations, and with double mutations. The accumulation of mutations in the gyrA gene was associated with an increase in fluoroquinolone resistance. Ofloxacin MICs for the majority of the strains with single and double mutations were 0.39 to 3.13 and 6.25 to 100 microg/ml, respectively. This study demonstrates a chronological increase in the percentage of not only highly fluoroquinolone-resistant strains, corresponding to those with double mutations in the gyrA gene, but also strains with moderately decreased susceptibilities to fluoroquinolones, corresponding to those with single mutations. This increase in the incidence of strains with a single mutation in the gyrA gene portends a further increase in the incidence of strains with clinically significant resistance to fluoroquinolones.

This article has been cited by other articles:

• Fricke, W. F., Wright, M. S., Lindell, A. H., Harkins, D. M., Baker-Austin, C., Ravel, J., Stepanauskas, R. (2008). Insights into the Environmental Resistance Gene Pool from the Genome Sequence of the Multidrug-Resistant Environmental Isolate Escherichia coli SMS-3-5. J. Bacteriol. 190: 6779-6794 [Abstract] [Full Text]  
• Smith, J. L., Drum, D. J. V., Dai, Y., Kim, J. M., Sanchez, S., Maurer, J. J., Hofacre, C. L., Lee, M. D. (2007). Impact of Antimicrobial Usage on Antimicrobial Resistance in Commensal Escherichia coli Strains Colonizing Broiler Chickens. Appl. Environ. Microbiol. 73: 1404-1414 [Abstract] [Full Text]  
• Alonso, R., Galimand, M., Courvalin, P. (2004). An extended PCR-RFLP assay for detection of parC, parE and gyrA mutations in fluoroquinolone-resistant Streptococcus pneumoniae. J Antimicrob Chemother 53: 682-683 [Full Text]  
• Kijima-Tanaka, M., Ishihara, K., Morioka, A., Kojima, A., Ohzono, T., Ogikubo, K., Takahashi, T., Tamura, Y. (2003). A national surveillance of antimicrobial resistance in Escherichia coli isolated from food-producing animals in Japan. J Antimicrob Chemother 51: 447-451 [Abstract] [Full Text]  
• McDonald, L. C., Chen, F.-J., Lo, H.-J., Yin, H.-C., Lu, P.-L., Huang, C.-H., Chen, P., Lauderdale, T.-L., Ho, M. (2001). Emergence of Reduced Susceptibility and Resistance to Fluoroquinolones in Escherichia coli in Taiwan and Contributions of Distinct Selective Pressures. Antimicrob. Agents Chemother. 45: 3084-3091 [Abstract] [Full Text]  
• Fluit, A. C., Visser, M. R., Schmitz, F.-J. (2001). Molecular Detection of Antimicrobial Resistance. Clin. Microbiol. Rev. 14: 836-871 [Abstract] [Full Text]  
• Giraud, E., Brisabois, A., Martel, J.-L., Chaslus-Dancla, E. (1999). Comparative Studies of Mutations in Animal Isolates and Experimental In Vitro- and In Vivo-Selected Mutants of Salmonella spp. Suggest a Counterselection of Highly Fluoroquinolone-Resistant Strains in the Field. Antimicrob. Agents Chemother. 43: 2131-2137 [Abstract] [Full Text]  
• Heurtin-Le Corre, C., Donnio, P.-Y., Perrin, M., Travert, M.-F., Avril, J.-L. (1999). Increasing Incidence and Comparison of Nalidixic Acid-Resistant Salmonella enterica subsp. enterica Serotype Typhimurium Isolates from Humans and Animals. J. Clin. Microbiol. 37: 266-269 [Abstract] [Full Text]