STUDY ON POLYMERASE CHAIN REACTION MOLECULAR DETECTION OF FIVE KINDS OF COMOM PATHOGENIC BACTERIA IN POLLUTED WATER

[Objective]To develop a multiplex PCR assay for simultaneously detecting Salmonella,Shigella,Pseudomonas aeruginosa,enterohemorrhagic Escherichia coli(EHEC)and Vibrio parahaemolyticus in polluted water,to provide experimental evidence for rapid diagnosis of these pathogenic bacterium infection.[Methods]Five pairs of specific primers were screened and designed to compose optimized system and conditions of multiplex PCR.[Results]The detection sensitivity of multiplex PCR were 102cfu,102cfu,102cfu,101cfu and 102cfu for EHEC,Shigella,Vibrio parahaemolyticus,Pseudomonas aeruginosa and Salmonella,respectively.The detection results of polluted water and natural water by multiplex PCR were consistent with that of traditional assay,and expected straps appeared to be clear and specific.Besides,it took multiplex PCR 6-8 hours to detect a sample.Compared with traditional assay,the detection time was shortened greatly,and the sensitivity improved obviously.So multiplex PCR method was more sensitive,specific and efficient,and the processing was rapid and simple.[Conclusions]The multiplex PCR method is suitable for detection of large sample,and it can be applied in food detection and environmental monitoring.