Detection and characterization of human rotavirus in tap water by multiplex RT-PCR

Abstract

More than a billion diarrheal cases occur each year among children below five years resulting in high morbidity 
and mortality. Rotaviruses, the single leading cause of diarrhea in children below five years, cause more than 
130 million episodes of severe diarrhea throughout the world. While treated water has been identified as a route 
of transmission, rotavirus in treated water has not been investigated in Ghana. Since the presence of 
rotaviruses in drinking water is unacceptable, a virological survey to detect and characterize rotavirus was 
conducted on tap water in the western part of Accra. Treated water samples were collected from five zones 
within the distribution network of Weija Water Works. Two litres of each sample were concentrated 4,000 fold 
and the viral particles extracted. Viral RNA was extracted from all concentrates using phenol/chloroform, 
purified with the RNaid®kit and reverse transcribed. The cDNA was amplified by semi-nested PCR using P and G 
genotype- specific primers and analyzed an agarose gel. Rotavirus was detected in 48.1% of samples. Nine 
rotavirus P-types and 12 G-types were detected. The detection pattern showed increased viral pollution with 
distance from the treatment plant and in areas with high human activity.