TaqMan PCR for Detection of Vibrio cholerae O1, O139, Non-O1, and Non-O139 in Pure Cultures, Raw Oysters, and Synthetic Seawater

摘要: Vibrio cholerae is recognized as a leading human waterborne pathogen. Traditional diagnostic testing for not always reliable, because this bacterium can enter viable but nonculturable state. Therefore, nucleic acid-based tests have emerged useful alternative to traditional enrichment testing. In article, TaqMan PCR assay presented quantitative detection of V. in pure cultures, oysters, and synthetic seawater. Primers probe were designed from the nonclassical hemolysin ( hly A) sequence strains. This was applied DNA 60 bacterial strains comprising 21 genera. The positive all tested negative other species tested. addition, none genera amplified with primers used study. results raw oysters spiked serotypes O1 O139 comparable those cultures. sensitivity range 6 8 CFU g −1 10 ml oyster seawater samples, respectively. total could be completed 3 h. Quantification cells linear over at least log units. primer set developed study rapid screening tool presence without prior isolation characterization bacteria by microbiological methods.