Detection of Enteroviruses and Mammalian Reoviruses by RT-PCR and Integrated Cell Culture-PCR in CPE-positive Surface Water Samples

Kim, Hee Jung; Ko, Hyun Ae; Kim, Sang Hyun

Abstract

The environmental water samples assayed by total culturable virus assay (TCVA) were evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and integrated cell culture-PCR (ICC-PCR) method for enteroviruses and reovirus since they are the usually detected virus groups by culture assays. The detection sensitivities of the TCVA, RT-PCR, and ICC-PCR were compared and the overall reliability of the detection was analyzed to confirm environmental samples for enteric viruses. A total of eight samples from different areas was analyzed by performing TCVA, RT-PCR, and ICC-PCR. Virus concentrations in surface water samples ranged from 1.03 to 47.3 most probable numbers of infectious units (MPN) per 100 liters. When primers specific for both enteroviruses and reoviruses were used in both RT-PCR and ICC-PCR, all the samples (100%) were positive for the viruses. Reoviruses were the most frequently detected ones among the samples. According to the sequence results of enteroviruses, five of the samples were contaminated by coxsackievirus type B3, and the rest by coxsackievirus type B6, echovirus type 30, or vaccine strain poliovirus type 3. It was observed that both enteroviruses and reoviruses were detected concurrently in all CPE-positive environmental water samples.