MOLECULAR EPIDEMIOLOGIC SURVEY OF ROTAVIRUSES IN SEWAGE BY REVERSE-TRANSCRIPTASE SEMINESTED PCR AND RESTRICTION-FRAGMENT-LENGTH-POLYMORPHISM ASSAY

Rotavirus double-stranded RNA was detected directly in sewage treatmen t plant samples over a 1-year period by reverse transcription followed by PCR amplification of the VP7 gene and Southern blot hybridization. The presence of naturally occurring rotaviruses was demonstrated in 4 2% of raw sewage samples and in 67% of treated effluent samples, Ampli fied viral sequences were analyzed bg restriction enzymes. Ten differe nt restriction profiles were characterized, most of which were found i n treated effluent samples. A mixture of restriction profiles was obse rved in 75% of contaminated effluent samples, The profiles were compar ed with those obtained from human rotavirus isolates involved in infec tions in children from the same area (six different profiles were dete cted), Five identical viral sequences were detected in both environmen tal and clinical samples, Restriction profiles sere also compared io p rofiles from known genomic sequences of human and animal viruses. Both human and animal origins of rotavirus contamination of water seemed l ikely.