Simple method of concentrating enteroviruses and hepatitis A virus from sewage and ocean water for rapid detection by reverse transcriptase-polymerase chain reaction

Key Result

RT-PCR was at least 500-fold more sensitive than cell culture for detecting enteroviruses and hepatitis A virus in sewage and ocean water, yielding results within 5 hours.

Key Points

• To establish a rapid, simple method for concentrating and detecting enteroviruses and hepatitis A virus in sewage and ocean water using RT-PCR.
• Concentrated sewage using Centriprep-100 and Centricon-100, and concentrated estuary and ocean water using vortex flow filtration and microconcentration.
• Amplified viral targets using RT-PCR with enterovirus or HAV capsid-specific primers, confirming products with a nonradioactive internal probe.
• Evaluated viral persistence at 4°C versus 25°C in seawater seeding experiments and compared RT-PCR performance directly against cell culture.
• RT-PCR achieved at least 500-fold higher sensitivity than cell culture and reduced detection turnaround time from 5 days to 3 weeks down to 5 hours.
• Seeding experiments demonstrated that hepatitis A virus persisted longer than poliovirus in seawater at 4°C, with both viruses showing extended persistence at 4°C compared to 25°C.