A one-step RT-PCR assay to detect and discriminate porcine reproductive and respiratory syndrome viruses in clinical specimens

Gene. 2013 Dec 1;531(2):199-204. doi: 10.1016/j.gene.2013.09.017. Epub 2013 Sep 11.

Abstract

Outbreaks of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) have led to large economic losses and, subsequently, have drawn great attention to its diagnosis and prevention. To facilitate rapid discrimination of HP-PRRSV from classical PRRSV (C-PRRSV), we developed a one-step RT-PCR assay. Primer specificities were evaluated with RNA extracted from 8 viral strains and our results revealed that the primers had a high specificity for PRRSV. The assay sensitivity was 25 copies/μL for both HP-PRRSV and C-PRRSV. A total of 929 serum samples were identified, of which 20.45% were HP-PRRSV-positive and 1.51% were C-PRRSV-positive, which was completely consistent with that of immunochromatochemistry and sequencing method. The proposed assay can detect the virus 2 days prior the onset of symptoms and it can be performed in 2h, thereby providing a rapid method to discriminate HP-PRRSV from C-PRRSV for the identification and prevention of PRRSV infections.

Keywords: CSFV; Highly pathogenic; JEV; Japanese encephalitis virus; One-step RT-PCR; PCV2; PPV; PRRSV; PRV; Porcine reproductive and respiratory syndrome virus; RV; Viral discrimination; classical swine fever virus; porcine circovirus type 2; porcine parvovirus; porcine reproductive and respiratory syndrome virus; pseudorabies virus; rotavirus.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA Viruses / genetics
  • Diagnosis, Differential
  • Porcine Reproductive and Respiratory Syndrome / blood
  • Porcine Reproductive and Respiratory Syndrome / diagnosis*
  • Porcine Reproductive and Respiratory Syndrome / genetics
  • Porcine Reproductive and Respiratory Syndrome / virology
  • Porcine respiratory and reproductive syndrome virus / genetics
  • Porcine respiratory and reproductive syndrome virus / isolation & purification*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Swine / blood
  • Swine / virology
  • Viruses / genetics