Detection of Enteroviruses

Jiménez-Clavero, Miguel-Angel; Ley, Victoria; Gómez, Nuria; Sáiz, Juan-Carlos

Enteroviruses are members of the Picornaviridae family and represent one of the most important water-transmitted pathogens. Detection of enteroviruses in water sources, or water-contaminated food, is a very valuable tool not only to prevent waterborne diseases but also to track down animal or human environmental viral pollution. Nowadays, molecular biology techniques allow the use of very sensitive and specific reverse-transcription polymerase chain reaction (RT-PCR) procedures to detect enteroviruses. In this chapter, using bovine enterovirus as a model, we describe procedures for enterovirus detection. Detailed descriptions of proper sample collection, storage, and processing, including methods for water concentration and solid sample extraction to obtain viral RNA, are outlined. Next, we describe methods for enterovirus detection based on virus isolation in appropriate cell culture. Finally, protocols for molecular detection of enterovirus are described, including procedures for conventional, nested, and real-time RT-PCR.