Multiplex PCR water testing kit for rapid, economic and simultaneous detection of Escherichia coli, Yersinia enterocolitica and Aeromonas hydrophila from drinking water

In recent years new or emerging pathogens have arisen as a problem in drinking water. Emerging human pathogens like Yersinia enterocolitica and Aeromonas hydrophila producing an impressive array of virulent factors causing both gastrointestinal and non-intestinal diseases in children and adults, have been isolated from drinking water. The conventional methods cannot predict the presence of these potentially enteropathogenic microorganisms in drinking water. To combat these limitations a multiplex PCR water testing kit has been designed, which can simultaneously detect Escherichia coli, Y. enterocolitica and A. hydrophila from water after pre-enrichment in a newly developed bacteriological water testing kit (BWTK). PCR can be performed directly by taking an aliquot of 200 mu l of water sample from non-potable BWTK in a PCR tube, heat treated at 95 degrees C for 10 min, and centrifuged at 10,000 rpm for 5 min; 5 mu l of supernatant was used for multiplex PCR. The high output and cost-effective multiplex PCR water testing kit developed in this study could provide a powerful supplement to conventional methods for more accurate risk assessment and monitoring of pathogenic bacteria in water.