Rapid quantification of infectious enterovirus from surface water in Bohai Bay, China using an integrated cell culture-qPCR assay.

To rapidly quantify infectious enteroviruses polluting the coastal seawaters, a newly developed integrated cell culture and reverse transcription quantitative PCR (ICC-RT-qPCR) assay was used to identify the contamination by enteroviruses in winter seawater samples of Bohai Bay, Tianjin, China. The gene copies of enteroviral 5'UTRs correlated to the initial inoculum numbers across the concentration range of 0.05-500 PFU mL(-1) (correlation coefficient (R(2)) was 0.9667). ICC-qPCR revealed that five of seven samples (70.4%) were positive for infectious enteroviruses. The concentration of enteroviruses was estimated at 0.2-21 PFU L(-1). The result demonstrated that the contamination of enteroviruses in this coastal area may constitute a potential public health risk. This study established a practical assay for widespread monitoring studies of aquatic environments for viral contamination and provided meaningful data for human waterborne viral risk assessment. Crown