SIMPLE METHOD OF CONCENTRATING ENTEROVIRUSES AND HEPATITIS-A VIRUS FROM SEWAGE AND OCEAN WATER FOR RAPID DETECTION BY REVERSE TRANSCRIPTASE-POLYMERASE CHAIN-REACTION

A rapid and simple method was developed to detect enteroviruses and hepatitis A virus (HAV) in sewage and ocean water. Sewage samples were concentrated by Centriprep-100 and Centricon-100 at 1,000 x g. Samples collected from estuary and near-shore surf zone ocean water in Southern California were concentrated by vortex, flow filtration and microconcentration. Reverse transcriptase-polymerase chain reaction (RT-PCR), with enterovirus primers or HAV capsid-specific primers, was used to detect enteroviruses or RAV in all concentrated samples. A nonradioactive internal probe was used to confirm the amplified products. Results of seeding experiments indicated that at 4-degrees-C, HAV was more persistent than poliovirus in seawater and both HAV and poliovirus persisted longer at 4-degrees-C than at 25-degrees-C. RT-PCR was at least 500-fold more sensitive than cell culture. Results were obtained within 5 h by RT-PCR, in contrast with the 5 days to 3 weeks required for cell culture.