Establishment and application of SYBR Green Ⅰ real-time fluorescence quantitative PCR method for chicken circle virus type 3
In this study,a standard plasmid based on GyV3 VP2 gene was constructed and used as a template to establish a standard curve.On the basis of optimizing the qPCR reaction system,the specificity,sensitivity and repeatability of this method were evaluated.The results revealed the method was only specific for GyV3 amplification,the minimum detection template concentration was 1.585 × 101 copies/μL,and the coefficient of variation in both intra-class and intra-group repea-tability tests was less than 1.0％.The qPCR method established in this study was used to detect 50 clinical samples,and the detection rate was 10％(5/50),higher than the conventional PCR detection rate of 8％(4/50).The results above indicate that the real-time fluorescent quantitative PCR method established in this study has high specificity,reproducibility,and sensitivity and can be utilized for quick clinical detection of GyV3.
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