DETECTION OF ADENOVIRUSES AND ENTEROVIRUSES IN POLLUTED WATERS BY NESTED PCR AMPLIFICATION

A procedure has been developed for the rapid detection of enteroviruse s and adenoviruses in environmental samples. Several systems for virus concentration and extraction of nucleic acid were tested by adding ad enovirus type 2 and poliovirus type 1 to different sewage samples. The most promising method for virus recovery involved the concentration o f viruses by centrifugation and elution of the virus pellets by treatm ent with 0.25 N glycine buffer, pH 9.5. Nucleic acid extraction by ads orption of RNA and DNA to silica particles was the most efficient. One aliquot of the extracted nucleic acids was used for a nested two-step PCR, with specific primers for all adenoviruses; and another aliquot was used to synthesize cDNA for a nested two-step PCR with specific pr imers for further detection of seeded polioviruses or all enteroviruse s in the river water and sewage samples. The specificity and sensitivi ty were evaluated, and 24 different enterovirus strains and the 47 hum an adenovirus serotypes were recognized by the primers used. The sensi tivity was estimated to be between 1 and 10 virus particles for each o f the species tested. Twenty-five samples of sewage and polluted river mater were analyzed and showed a much higher number of positive isola tes by nested PCR than by tissue culture analysis. The PCR-based detec tion of enteroviruses and adenoviruses shows good results as an indica tor of possible viral contamination in environmental wastewater.