Detection of porcine enteroviruses by nRT-PCR: differentiation of CPE groups I-III with specific primer sets

Porcine enteroviruses (PEV) comprising at least 13 serotypes grouped into t hree species are described as causative agents of neurological disorders, f ertility disorders, and dermal lesions of swine. Despite their well-documen ted acid stability, enteric infection route, and similarity of clinical sym ptoms, most of the porcine enterovirus (PEV) serotypes are set apart from t he genus Enterovirus of the Picornaviridae. Hence, PCR procedures used comm only to detect enteroviruses are not applicable to epizootic relevant PEV s erotypes. A nested RT-PCR protocol is described now suited to detect all kn own porcine enterovirus serotypes using three sets of primer pairs. These p rimer pairs were designed to amplify either highly conserved sequences of t he 5'-nontranslated region (5'-NTR or the polymerase gene region of the rel evant virus species. All 13 acknowledged serotypes of three PEV species and several field isolates of clinical specimens were detectable. The specific ity of the PCR procedure is supported by the observation that RT-PCR-positi ve field isolates coincide with serological PEV classification. PEV PCR is more rapid and less laborious than the time-consuming virus isolation by ti ssue culture techniques over several passages and serotyping. Because other viruses such as classical swine fever virus, pseudorabies virus, porcine p arvovirus, swine vesicular disease virus, and foot-and-mouth disease virus may cause diseases with similar clinical symptoms, PCR detection of all PEV s closes a diagnostic gap and offers the opportunity to use comprehensive P CR procedures for the diagnosis of all relevant viruses causing such sympto ms. (C) 2000 Elsevier Science B.V. All rights reserved.