Comparative detection of enteric viruses in wastewaters, sediments and oysters by reverse transcription-PCR and cell culture

The work presented here examines the utility of reverse transcription-PCR (RT-PCR) assays for monitoring enteric viruses contaminating wastewaters, sediments and shellfish. Sampling occurred over a 12 month period from and around a large cosmopolitan sewage treatment facility in Auckland, New Zealand. Viruses were concentrated using primary polyethylene glycol 6000 (PEG 6000) precipitation and recently developed secondary concentration and purification techniques as preliminary steps to analysis by plaque assay or RT-PCR for enteroviruses, rotaviruses and hepatitis A virus (HAV). Enteroviruses were isolated by plaque assay from each of the different sample types at various points during the year. All three groups of viruses were detected by the PCR in different sample types and at various time points. The results demonstrated that RT-PCR was most useful when examining samples for viruses routinely difficult to identify, namely rotaviruses and HAV.