Development and application of real-time polymerase chain reaction to detect Vibrio cholerae O1 and O139 in river water

• Published in: Zhōnghuá liúxíngbìng zázhì Vol. 28; no. 8; p. 768
• Main Authors: Wang, Xiao-Mei, Wang, Duo-Chun, Tan, Hai-Ling, Zhong, Hao-Jie, Chen, Jing-Diao, Li, Bai-Sheng, Ke, Chang-Wen, Yan, Mei-Ying, Zhang, Jing, Kan, Biao
• Format: Journal Article
• Language: Chinese
• Published: China 01.08.2007
• Subjects: Environmental Monitoring - methods; Genes, Bacterial; Polymerase Chain Reaction - methods; Reproducibility of Results; Rivers - microbiology; Sensitivity and Specificity; Vibrio cholerae O1 - isolation & purification; Vibrio cholerae O139 - isolation & purification
• ISSN: 0254-6450

To develop a real-time SYBR Green polymerase chain reaction (PCR) for detection of Vibrio cholerae serogroups O1 and O139, and to evaluate its reliability through detection of estuary water samples. O antigen rfb genes specific for O1 and O139 were used for the design of PCR primers. The real-time SYBR Green PCR system in detecting O1 and O139 specific rfb genes in one tube was developed, and its sensitivity, specificity and reproducibility were evaluated. The ability of the real-time PCR in detection of estuary water samples was compared with the routine PCR and bacteria isolation. The amplification of O1 or O139 specific target gene could be detected according to the melt curve temperature of amplicons. No amplification was observed in the templates of other 10 non-cholerae vibrios. When comparing to the real-time PCR to bacteria isolation in detection of 524 estuary water samples, it showed high sensitivity, plus also positive in real-time PCR detection among all the samples in which bacteria of O1 or O139