Novel one step real time RT-PCR Assay for the detection of Enterovirus

nteroviruses (EV) are the leading cause of aseptic meningitis in pediatric and adult populations and can be associated with severe disease such as myocarditis, encephalitis, and paralytic poliomyelitis. RT-PCR has rapidly become the diagnostic methodology of choice due to its sensitivity and rapid turn-around-time allowing significant improvement in patient care and management. Most molecular assays target the highly conserved regions within the 5' nontranslated region (NTR) described by Rotbart el al. We describe the development of a novel real time assay that amplifies and detects a conserved region upstream of the Rotbart amplicon utilizing primers and an Eclipse probe. As a further enhancement, an RNA internal control (IC) is integrated into the reaction. The analytical sensitivity was determined and a comparison to the Chemicon Oligodetect Panenterovirus kit was made during the 2005 enterovirus season.