Development of a real-time PCR assay for detection of porcine circovirus type 1

The objective of this study was to develop a real-time quantitative fluorescence PCR assay for quantitative detection of porcine circovirus type 1(PCV1).A pair of primers specific to the PCV1 gene was designed to construct a recombinant plasmid,and the positive recombinant plasmid was diluted serially as templates to establish a real-time PCR assay based on SYBR GreenⅠ for quantitative detection of PCV1.Meanwhile,the corresponding standard curve was generated,and the specificity,sensitivity and reporducibility of this assay were determined.The coefficient of determination for the developed standard curve was 0.999,which indicated an excellent linear relationship.The assay had an accurate detectable limit of 320copies/μL,and its coefficient of variation was less than 2%in the reproducible assays.No amplification was detected by this method from unrelated swine virus samples,including PCV2,porcine parvovirus,pseudorabies virus,porcine reproductive and respiratory syndrome virus and classical swine fever virus.The detectable rate of several clinical samples with this assay was higher than that of conventional PCR assay.The results indicated that this real-time PCR assay was of high specificity,sensitivity and reproducibility,and could be used for the detection and quantitative analysis of PCV1.