Development of TaqMan fluorescent quantitative PCR for detection of porcine circovirus type 2

A pair of primers and a probe were designed according to the conserved sequence of ORF1 gene of porcine circovirus type 2(PCV2)and the recombinant plasmid containing the complete genome sequence was constructed as a standard control,then a fluorescent quantitative real-time PCR(FqRT-PCR) method was developed by optimizing reaction conditions and tests of specificity,sensitivity and reproducibility were carried out.In results,the developed method could specifically detect PCV2,and had no cross- reaction with porcine circovirus type 1and other viruses.The sensitivity of this method was proved to be 1×102 copies/μL,which was 100fold better than the traditional PCR,and the coefficient of variation value was less than 5%.Among 94samples,the positive rate was 51.06%,detected by FqRT-PCR,38.30%by the traditonal PCR.The results showed that the developed method had the advantages of rapidity,specificity,reproducibility,quantitativity and sensitivity,and it was able to be applied to clinical diagnosis.