Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10030473 | Journal of Virological Methods | 2018 | 29 Pages |
Abstract
The aim of this study was to develop a multiple PCR assay based on the suspension array system for the simultaneous detection of respiratory and reproductive pathogens in swine. Pseudorabies virus (PRV), Japanese encephalitis virus (JEV), classic swine fever virus (CFSV), African swine fever virus (ASFV), porcine circovirus type 2 (PCV-2), porcine reproductive and respiratory syndrome virus (PRRSV) and porcine parvovirus (PPV) are the major respiratory and reproductive viral pathogens in pig farms. Seven pairs of specific primers and probes were designed, and the multiple PCR was performed, with the PCR products hybridized to beads coupled to probes, which were then detected by Bio-Plex suspension array system. The limit of detection, specificity and repeatability of this method was determined. The assay was further tested using 137 clinical samples, and the results were compared with conventional PCR to evaluate the ability of the method to diagnose porcine viruses. The results showed that the assay had a high degree of specificity and repeatability, and the simultaneous detection limit for the seven viruses reached 103 copies/μL. Forty-nine of the clinical samples tested positive for at least one of the viruses, the principal viral infections in the clinical samples were PCV-2 and PRRSV. The suspension method represented a rapid, specific and high-throughput tool for single or mixed detection of the seven porcine viruses simultaneously, and has great significance for the development of liquid chip techniques for the diagnosis of diseases in animals.
Keywords
PRVSA–PETris–EDTAASFVTMACPasteurella multocidaHaemophilus parasuisPEDVFMDVTGEVRCSHPSSDSMultiplex PCRPPVEDCMFIPRRSVTetramethylammonium chlorideSuspension arraystandard deviationMolecular diagnosticJEVsodium dodecyl sulfatemedian fluorescence intensityCoefficient of VariationMeSPorcine circovirus type 2Pseudorabies virusJapanese encephalitis virusPorcine epidemic diarrhea virusAfrican swine fever virusPorcine reproductive and respiratory syndrome virusTransmissible gastroenteritis virusPorcine parvovirus
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Authors
Lu Xiao, Yin Wang, Runmin Kang, Xulong Wu, Hua Lin, Yonggang Ye, Jifeng Yu, Jianqiang Ye, Jing Xie, Ye Cao, Yong Wei, Dangjin Liao, Meng Pan, Yi Lin, Zhuojian Dai, Xingyu Li,