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

J Virol Methods. 2014 Mar:198:69-74. doi: 10.1016/j.jviromet.2013.12.003. Epub 2013 Dec 21.

Abstract

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 103copies/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.

Keywords: Detection; Minor groove binder probe; Porcine Sapelovirus; Real-time PCR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5' Untranslated Regions / genetics
  • Animals
  • DNA Primers / genetics
  • RNA Viruses / genetics*
  • RNA, Viral / genetics*
  • Real-Time Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Swine
  • Swine Diseases / virology*

Substances

  • 5' Untranslated Regions
  • DNA Primers
  • RNA, Viral