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J Med Microbiol 56 (2007), 346-351; DOI: 10.1099/jmm.0.46655-0
© 2007 Society for General Microbiology
ISSN 1473-5644

Multiplex PCR for detection of antibiotic resistance genes and the SXT element: application in the characterization of Vibrio cholerae

Dhanya Ramachandran1, R. Bhanumathi1,{dagger} and Durg V. Singh2

1 Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram-695 014, India

2 Institute of Life Sciences, Nalco Square, Bhubaneswar-751 023, India

Correspondence
Durg V. Singh
durg_singh{at}yahoo.co.in

Received 3 April 2006
Accepted 26 October 2006

This study describes a multiplex PCR assay for the detection of antibiotic resistance genes and the SXT element in Vibrio cholerae. Conditions were optimized to amplify fragments of sulII (encoding sulfamethoxazole resistance), dfrA1 (O1-specific trimethoprim resistance), dfr18 (O139-specific trimethoprim resistance), strB (streptomycin B resistance) and the SXT element simultaneously in one PCR. This multiplex PCR was evaluated on 142 V. cholerae isolates and the results correlated with the phenotypic antibiotic data obtained using a disc diffusion assay and a colony blot assay. Thus this one-step PCR can be used as a simple, rapid and accurate method for identification of antibiotic resistance profiles and could be used for the surveillance of the spread of antibiotic resistance determinants in epidemiological and environmental studies.

Abbreviations: Cm, chloramphenicol; Fr, furazolidone; Str, streptomycin; Sul, sulfamethoxazole; Tmp, trimethoprim.

{dagger}Present address: Patent Office, Chennai Branch, IPR Building, GST Road, Chennai-600 032, India.






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