猪圆环病毒I型荧光定量PCR检测方法的建立

Establishment of a fluorescent quantitative PCR assay using TaqMan probe for PCV1 detection

以定量的10倍系列稀释的质量pMD-ORF2为标准品进行荧光定量PCR扩增，建立了PCV1检测的荧光定量PCR方法。结果表明该方法检测灵敏度可达1．0×10^4拷贝／μL，线性范围为10^10～10^1；对起始浓度为1．0×10^2、1．0×10^6、1．0×10^6拷贝／μL的标准品的最终实际测得值分别为1．001×10^7、0．869×10^6和0．988×10^5拷贝／μL，变异系数分别为4．758％、1．865％和3．836％，说明此方法具有良好的准确性和重现性。对阳性组织病料的检测表明该方法的检测灵敏度高出常规PCR，与套式PCR（nPCR）具有相近的灵敏度。

The FQ-PCR assay was carried out by quantitative concentration of serial 10 fold dilutions of pMD-ORF2 DAN by optimizing circulation parameters. In result, the sensitivity could reach 1.0×10^1 copy/μL and the linear relation was excellent. Meanwhile, the final measure value on initiative concentration of 10 ×10^7 ,10×10^6 ,10 ×10^5 copies/μL DAN was 1. 001 ×10^7 ,0. 869×10^6 and 0. 988×10^4 copies/μL and the coefficient of variation was 4.758% ,1.865% and 3. 836%, respectively. Furthermore, the sensitivity of FQ-PCR was roughly equal to that of nPCR and was prior to that of PCR by detecting the positive field samples.