Multiplex PCR assay for simultaneous detection and differentiation of , and spp. in the municipality-supplied drinking water.

BACKGROUND

The contamination with , , and spp. in drinking water is the most prevalent in Indian subcontinent, but often difficult to detect all these pathogens from the drinking water.

MATERIALS AND METHODS

A multiplex polymerase chain reaction (mPCR) method was developed to detect contamination of municipality-supplied drinking water with , , and spp. The primers were designed to target small subunit of 16S rRNA type gene of and , and invasive A gene of . The optimized mPCR assay was applied on 158 municipality-supplied drinking water samples collected from Delhi.

RESULTS

Out of total 158 water samples, 89 (56.32%) were found positive for the targeted pathogens by mPCR while conventional methods could be detected only in 11 (6.96%) samples. The mPCR assay showed 100% sensitivity and specificity for these pathogens in comparison with culture and microscopic detection. Of the 89 mPCR-positive samples, , , and spp. were present in 35 (22.15%), 26 (16.45%), and 28 (17.72%), respectively. Nine (5.69%) samples were positive for both and , 10 (6.32%) were positive for and spp., and 8 (5.06%) had spp. and . Nonetheless, 3 (1.89%) samples were positive for all three pathogens.

CONCLUSIONS

The present assay is an alternative to conventional methods to serve as highly sensitive, specific, and economical means for water quality surveillance to detect the outbreak caused by , , and spp. pathogens.