Ultrafiltration and reverse transcription-polymerase chain reaction: An efficient process for poliovirus, rotavirus and hepatitis A virus detection in water

A process to concentrate viruses from water associated with a rapid and sen sitive viral assay was evaluated with water samples experimentally seeded w ith a single virus or a virus mixture [poliovirus, rotavirus, hepatitis A v irus (HAV)]. Tangential ultrafiltration was used for virus concentration. R everse transcription-polymerase chain reaction was tested for virus detecti on and its sensitivity compared to that of cell culture. We recovered the t hree viruses from experimentally contaminated samples, and detected viral i nfectivity or RNA with low inputs: 1 TCID50 L-1 for cell culture vs 10(-3) TCID50 L-1 with the RT-PCR assay in the case of poliovirus, 1 TCID50 L-1 in the case of rotavirus whatever the technique, and RT-PCR allowed detection of HAV RNA till at least 1 TCID50 L-1. Ninety tapwater samples were also t ested for the presence of enterovirus and rotavirus. Five tapwater samples were positive for the RT-PCR assay only: one for poliovirus and four for ro tavirus. This procedure allows a control of the virological quality of wate r. (C) 2000 Elsevier Science Ltd. All rights reserved.