Establishment and Application of TaqMan Fluorescence Quantitative PCR for Detection of Porcine Circovirus Type 2

The probes and primers were designed according to the nucleotide sequence of porcine circovirus type 2(PCV2) available in GenBank, and real-time TaqMan fluorescence quantitative PCR for detection of PCV2 was established successfully.Clinical diagnosis of PCV2 samples and the 30 suspected samples were detected by using the established quantitative PCR which was compared with that of routine PCR.The results indicated that the established quantitative PCR assay could detect 4.5103 copysL-1 of plasmid DNA.Sensitivity and positive rate for clinical samples of TaqMan fluorescent quantitative PCR were higher than routine PCR, and its sensitivity was 10 times higher than that of the routine PCR.The 30 suspected samples were detected by TaqMan fluorescence quantitative PCR and routine PCR, respectively, the positive detection rate were 66.7% and 56.7%, the coincidence rate was 90%.Classical swine fever virus(CSFV), porcine reproductive and respiratory syndrome virus(PRRSV) and pseudorabies virus(PRV) were detected by using the established quantitative PCR and the result indicated that CSFV, PRRS and PRV were negative, and had no cross reaction.The real-time TaqMan fluorescence quantitative PCR assay which is specific,sensitive and accurate can be used for the clinical diagnosis and epidemiological investigation of PCV2.