title: Triplex reverse transcription-PCR for detecting viable toxigenic Vibrio cholerae in water samples in Thailand.
abstract: Abstract. Detection of toxigenic Vibrio cholerae O1/O139 in aquatic environment is difficult to achieve using the culture method. For direct detection of viable toxigenic V. cholerae in aquatic environment, we developed a triplex reverse transcription (RT)-PCR, targeting genes for the outer membrane protein (ompW), cholera toxin A (ctxA) and toxin-coregulated pilli (tcpA) and compared the assay with the culture method. After enrichment of the bacteria-containing filters in alkaline peptone water for 6 hours, the sensitivity of triplex RT-PCR for detecting V. cholerae was 7 cfu/ml. Of the 80 environmental water samples collected from various regions in Northeast Thailand, triplex RT-PCR detected 15 toxigenic and 20 non-toxigenic V. cholerae, whereas the culture method detected only 3 toxigenic V. cholerae--containing water samples. These results show that this triplex RT-PCR method could be used as an alternative tool for rapid and sensitive identification of viable toxigenic V cholerae in environmental water samples.
authors: Boonnapa
authors: B
authors: Kanoktippornchai
authors: Chariya
authors: C
authors: Chomvarin
authors: Chulapan
authors: C
authors: Engchanil
authors: Warawan
authors: W
authors: Wongboot
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013785:Thailand; D014734:Vibrio cholerae / Q000302:isolation & purification*; D014871:Water Microbiology