title: Establishment and application of a multiplex PCR for rapid and simultaneous detection of six viruses in swine.
abstract: A multiplex PCR assay was developed and evaluated subsequently for its effectiveness in simultaneously detecting mixed viral infections of swine. Specific primers were designed and used for testing the six swine viruses: three DNA viruses, including pseudorabies virus (PRV), porcine parvovirus (PPV), and porcine circovirus type 2 (PCV2); three common RNA viruses, including porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), and Japanese encephalitis virus (JEV). This technique has shown to be highly sensitive in that the minimum detection amounts of nucleic acids from PRV, PPV, PCV2, PRRSV, CSFV, and JEV were 6.6, 96, 12.9, 10.5, 51, and 46 pg, respectively. It also was effective for detecting one or multiple viruses in the specimens, such as the lungs, spleens, lymph nodes, and tonsils collected from clinically ill pigs. The multiplex PCR method can detect simultaneously not only infection of the six viruses, but also other swine DNA and RNA viruses. Given its rapidity, specificity, and sensitivity, the multiplex PCR is a useful tool for diagnosing clinically the mixed infections of swine DNA and RNA viruses.
authors.affiliation: College of Animal Sciences, Guizhou University, Guiyang 550025, China; Key Laboratory of Animal Epidemic Disease and Veterinary Public Health in Guizhou Province, Guiyang 550025, China. Electronic address: as.zyzeng@gzu.edu.cn.
authors.forename: Zhiyong
authors.initials: Z
authors.lastname: Zeng
authors.affiliation: College of Animal Sciences, Guizhou University, Guiyang 550025, China.
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Liu
authors.affiliation: College of Animal Sciences, Guizhou University, Guiyang 550025, China.
authors.forename: Weicheng
authors.initials: W
authors.lastname: Wang
authors.affiliation: College of Animal Sciences, Guizhou University, Guiyang 550025, China.
authors.forename: Deyuan
authors.initials: D
authors.lastname: Tang
authors.affiliation: Teaching and Experimental Farms, Guizhou University, Guiyang 550025, China.
authors.forename: Haiying
authors.initials: H
authors.lastname: Liang
authors.affiliation: College of Animal Sciences, Guizhou University, Guiyang 550025, China.
authors.forename: Zhao
authors.initials: Z
authors.lastname: Liu
mesh_terms: D000818:Animals; D060085:Coinfection / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology; D013997:Time Factors; D014730:Veterinary Medicine / Q000379:methods*; D014777:Virus Diseases / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D014780:Viruses / Q000235:genetics / Q000302:isolation & purification*
keywords: CSFV; JEV; Multiplex PCR; PCV2; PRRSV; PRV