title: A one-step RT-PCR assay to detect and discriminate porcine reproductive and respiratory syndrome viruses in clinical specimens.
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.
authors.affiliation: Hubei Key Laboratory of Animal Embryo and Molecular Breeding, Institute of Animal Husbandry and Veterinary, Hubei Academy of Agricultural Sciences, Wuhan 430064, PR China. Electronic address: keliy6@126.com.
authors.forename: Keli
authors.initials: K
authors.lastname: Yang
authors.forename: Yanhe
authors.initials: Y
authors.lastname: Li
authors.forename: Zhengying
authors.initials: Z
authors.lastname: Duan
authors.forename: Rui
authors.initials: R
authors.lastname: Guo
authors.forename: Zewen
authors.initials: Z
authors.lastname: Liu
authors.forename: Danna
authors.initials: D
authors.lastname: Zhou
authors.forename: Fangyan
authors.initials: F
authors.lastname: Yuan
authors.forename: Yongxiang
authors.initials: Y
authors.lastname: Tian
mesh_terms: D000818:Animals; D004267:DNA Viruses / Q000235:genetics; D003937:Diagnosis, Differential; D019318:Porcine Reproductive and Respiratory Syndrome / Q000097:blood / Q000175:diagnosis* / Q000235:genetics / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013552:Swine / Q000097:blood / Q000821:virology; D014780:Viruses / Q000235:genetics
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