COMPARISON OF CELL-CULTURE AND A POLIOVIRUS GENE PROBE ASSAY FOR THE DETECTION OF ENTEROVIRUSES IN ENVIRONMENTAL WATER SAMPLES

Nucleic acid hybridization provides a rapid non-cell culture method fo r the detection of enteric viruses in water. The purpose of this work was to compare the detection of naturally occurring enteroviruses by c ell culture with their detection by a poliovirus gene probe in various types of water samples. Samples of activated sludge effluent, tertiar y treated wastewater (activated sludge, filtration and passage through reverse osmosis), ground water, surface water and tidal river water w ere processed through 1 MDS Virozorb filters to concentrate any natura lly occurring virus. Viruses were eluted from the filters with pH 9.5 beef extract and reduced in volume by flocculation to 20-30 ml. These concentrates were then assayed in the BGM cell line by the cytopathoge nic effects (CPE) method and by a poliovirus cDNA probe (base pairs 11 5-7440) labeled with P-32. A total of 233 samples were assayed in this manner. In slightly more than 93% of the samples gene probe and cell culture yielded the same results. Of these samples 36 were positive by gene probe and 28 by cell culture assay. Positive samples for gene pr obe were confirmed by treatment with NaOH or RNAse and then reprobed. Samples demonstrating CPE upon primary passage were confirmed positive by subsequent passage of cell lysate on a new monolayer of BGM cells. Ten samples were positive by gene probe and negative by cell culture, and 4 samples were negative by gene probe and positive by cell cultur e.