DETECTION OF ADENOVIRUS AND ENTEROVIRUS BY PCR AMPLIFICATION IN POLLUTED WATERS

Several systems for virus recovery from environmental samples and extr action of nucleic acid were tested by adding adenovirus 2 and poliovir us 1 to different sewage samples. The most promising method involved: concentration of viruses by centrifugation, and elution of the pellete d viruses by treatment with 0.25 N glycine buffer pH 9.5. The nucleic acids were extracted by adsorption of RNA and DNA to silica particles. One aliquot was directly used for a two-step PCR in a nested fashion, with specific primers for all adenoviruses; the other aliquot was use d to synthesize cDNA and a nested two-step PCR with specific primers f or enteroviruses. The specificity and sensitivity of the selected prim ers were evaluated, the 47 human adenovirus serotypes were identified and 24 different enterovirus strains were-recognized. The sensitivity of the nucleic acid extraction, cDNA synthesis and nested PCR amplific ation was estimated to be between 1 and 10 viral particles. Sewage and polluted river samples were analyzed showing, as expected, a much hig her number of positive samples by the method described than by tissue culture analysis, a high prevalence of hepatitis A virus in sewage and the adenoviruses as the most commonly detected virus in the environme ntal samples analyzed.