Multiplex PCR for detection of antibiotic resistance genes and the SXT element: application in the characterization of Vibrio cholerae.
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.
Ramachandran
Dhanya
D
Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram-695 014, India.
Bhanumathi
R
R
Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram-695 014, India.
Singh
Durg V
DV
Institute of Life Sciences, Nalco Square, Bhubaneswar-751 023, India.
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*
D004251:DNA Transposable Elements; D004269:DNA, Bacterial