Development and Application of a Real-time Fluorescence Quantitative PCR Detection Method for Mink Circovirus
Specific primers were designed according to the Mink circovirus(MiCV)cap gene sequence and synthesized for development of a SYBR Green Ⅱ real-time fluorescence quantitative PCR.The results showed that the method had a good linear relationship in the template concentration range of 1.0×106 copies/μL to 1.0×101 copies/μL and the correlation coefficient was 0.9983.This detection method also had no amplifications for AMDV,PCV2,MEV,CDV and PRV.The intra-assay and inter-assay CV were less than 2％.The lower limit of recombinant positive plasmid was 1.0×101 copies/μL,which was 100 times more sensitive than the conventional PCR.In addition,the lower limit of detection for DNA of positive samples was 2.38×10-2 pg/μL,which was 1000 times more sensitive than the conventional PCR.Samples were collected from minks,foxes,raccoon dogs and environmental resources on some fur animal farms in Jilin province and tested using this method.The results showed that the positive rates were 57.23％for minks,48.42％for foxes,39.71％for racoon dogs and 68.48％for environmental resources,respectively.The above results indicated that the real-time fluorescence quantitative PCR method developed in this study had good sensitivity,specificity and repeatability,and provided technical support for the diagnosis and epidemiological investigation of MiCV.
Mink circovirusCap geneSYBR green Ⅱ real-time fluorescence quantitative PCR
国家科技期刊平台
NSTL
万方数据
维普
