Detection of Norovirus in Water by RT-PCR

Abstract:

Norovirus is recognized as one of the major cause agents of foodborne or waterborne non-bacterial gastroenteritis. The development of RT-PCR for detection of norovirus in fecal samples, and then the potential usefulness of the assay were confirmed for detection of water samples which were contaminated by norovirus in experiment circumstance. The specificity and sensitivity were also evaluated in the assay. The anticipated band of 327bp were obtained when the primer set of JV12/JV13 were used, which targeting RNA dependent polymerase. The specific amplicons were further confirmed by southern hybridization and the same results obtained after many repeats. The detection limits were 50pg/mL in fecal samples, and 200pg/mL in artificially contaminated water samples. In a total of 42 experimentally contaminated water samples, 38 samples were positive for norovirus and 4 pond water samples were negative. The results may stem from unrecovery of norovirus and the inhibitors in these water samples. The assay developed in this study can be applied to screening norovirus in water, and can attribute to the control of water quality.