Detection of all seven serotypes of foot-and-mouth disease virus by real-time, fluorogenic reverse transcription polymerase chain reaction assay

Abstract

A fluorogenic RT-PCR (5′-nuclease probe-based) assay using a primer/probe set designed from the internal ribosomal entry site region of the virus genome was developed for the specific detection of all seven serotypes of foot-and-mouth disease (FMD) virus in epithelial suspensions and cell culture virus preparations. The reverse transcription polymerase chain reaction (RT-PCR) specifically detected FMD virus in sample submissions from the UK 2001 FMD outbreak with greater sensitivity than our conventional RT-PCR procedure and our routine diagnostic procedures of ELISA and virus isolation in cell culture. The fluorogenic RT-PCR provides relatively fast results, enables a quantitative assessment to be made of virus amounts and can handle more samples and/or replicates of samples in a single assay than the conventional RT-PCR procedure. Therefore it is seen as a valuable tool to complement the routine diagnostic procedures for FMD virus diagnosis.