Development of a minor groove binder assay for real-time PCR detection of porcine Sapelovirus

A 5′ conjugated minor groove binder (MGB) probe real-time PCR assay was developed in this study for porcine sapelovirus (PSV) detection and quantitation. Two primers and a MGB probe for the 5′ untranslated region (UTR) gene were designed. The assay was capable of detecting about 103 copies/μl of standard template per reaction. Moreover, it does not detect any of the other RNA viruses that cause diarrhea disease in pigs. The coefficients of variation of intra- and inter-assay reproducibility were both lower than 2%. In 73 field fecal samples, PSV was detected in 46 samples using real-time PCR assay and only 32 samples with a conventional PCR assay. Therefore, the availability of this assay will facilitate further studies on the epidemiology of PSV infection and its role in swine disease.