title: Multiplex PCR for detection of antibiotic resistance genes and the SXT element: application in the characterization of Vibrio cholerae.
abstract: 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.
authors.affiliation: Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram-695 014, India.
authors.forename: Dhanya
authors.initials: D
authors.lastname: Ramachandran
authors.affiliation: Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram-695 014, India.
authors.forename: R
authors.initials: R
authors.lastname: Bhanumathi
authors.affiliation: Institute of Life Sciences, Nalco Square, Bhubaneswar-751 023, India.
authors.forename: Durg V
authors.initials: DV
authors.lastname: Singh
mesh_terms: D002771:Cholera / Q000382:microbiology; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D005798:Genes, Bacterial; D005838:Genotype; D006801:Humans; D007194:India; D008826:Microbial Sensitivity Tests; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D013223:Statistics as Topic; D014734:Vibrio cholerae / Q000187:drug effects / Q000235:genetics*
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial