Development and Application of a Specific Fluorescent Quantitative PCR Assay for the Detection of Porcine Circovirus Type 2

A fluorescent quantitative PCR based on SYBR Green I was developed for the detection of porcine circovirus type 2.A pair of primers were designed specific for PCV 2 according to the published nucleotide sequence(GenBank accession No.HQ148879).Using PCR assay,a 702 bp region of the PCV 2 ORF2 gene was cloned into pVAX1 vector.Serial dilutions of plasmid PCV 2 were used to quantify the virus genomic copy number,and served as standard curve in the fluorescent quantitative PCR for development of a specific assay for the detection of porcine circovirus type 2.The results showed that the fluorescence quantitative PCR assay was able to detect PCV 2.The sensibility of this assay attained 10 copies of plasmid DNA,which was 100 times higher than routine PCR.The C.V.of the intra or inter was no more than 2.5%,indicating that the developed fluorescence quantitative PCR is a stable assay.The clinical detection showed the development of the method achieved the effct of rapid diagnosis and quantitative detection of PCV 2.