Rapid identification of three waterborne pathogens in water samples by multiplex touchdown PCR

摘要

The purpose of this study was to develop a multiplex touchdown PCR to identify three waterborne pathogens in water samples. Specific primers based on VP1332, inv and ipaH gene sequences were designed for V. parahaemolyticus, Y. enterocolitca and Shigella, respectively. By optimizing reaction conditions, assays obtained high specifity, sensitivity and repeatability. The detection limit can be as low as 0.005ng/μl V. parahaemolyticus and 0.5ng/μl Y. enterocolitca and 0.005ng/μl of Shigella respectively. The detection process was about 3～4 hours, saving a lot of time compared with microbiological culture method that required 2～3 days. The system could be useful for the detection of multiple pathogens in water samples to protect public health.