Concentration, Detection and Identification of Infectious Enteroviruses in Sewage

Abstract

This work presents a new approach for the molecular 'serotyping' of enteroviruses concentrated and isolated from sewage. Viruses were adsorbed to cellulose nitrate membrane filters, isolated from the membrane filter using the VIRADEN method and identified by RT-PCR, followed by 57-UTR RFLP analysis and partial sequencing of the VP1 protein coding region. All 42 isolates belonged to the same genetic sub-cluster of non-polio enteroviruses. Partial VP1 sequencing revealed that isolates belonged to serotypes CBV4 (33 isolates, 79%), CBV1 (8 isolates, 19%) and Echovirus7 (1 isolate, 2%). The methodology presented here is highly promising for environmental surveillance of enterovirus circulation and epidemiology. 57-UTR RFLP analysis with Hpall is useful for initial subclassification, while partial VP1 sequencing is efficient for molecular 'serotyping'. © 2006 Wiley-VCH Verlag GmbH & Co. KGaA.