• Biomedical Science Letters
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• v.29 no.2
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• pp.81-87
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• 2023

Pan-Enterovirus (Pan-EV) infects millions of children and infants worldwide every year. As severe infections have recently been reported, the need for monitoring has consequently intensified. Pan-EV is a categorical name for waterborne enteroviruses belonging to the Picornaviridae family, and includes a wide range of pathogens including Coxsackievirus (CoxV), Echovirus (EcoV) and Enterovirus (EV). In this study, we proposed an optimal RT-nested PCR method for diagnosis of various types of Pan-EV in an aquatic environment and developed a positive control. Considering detection sensitivity, specific reaction, and final identification, one condition capable of amplifying 478 bp among the four candidates in the 1^st round PCR (RT-PCR) and one condition in the 2^nd round PCR (nested PCR) were selected. Through the detection of nucleic acids extracted from 123 groundwater samples and the detection sensitivity test based on artificial spiking in the sample, the methods are optimal for non-disinfected water samples such as groundwater. We developed a positive control for Pan-EV detection that can be amplified to different sizes under the two conditions. Accuracy could be further improved by testing for contamination from the control group. The method proposed in this study and the positive control developed are expected to be used in monitoring Pan-EV in aquatic environments including groundwater through future research using more samples.