用实时荧光定量PCR技术检测奶牛瘤胃纤维降解菌相对定量方法的研究

The Ruminal Fibrolytic Bacteria from Lactating Cows Revealed by Relative Quantification Real-time PCR

为了选择适合反刍动物瘤胃纤维降解菌的实时定量PCR相对定量方法,选用三头安装有永久性瘤胃瘘管的荷斯坦奶牛,针对奶牛瘤胃主要纤维降解菌,使用三种实时荧光定量PCR相对定量方法进行比较研究。结果表明:在相同条件下,对于同一个菌株基因的相对定量结果来说,SCF法和Pfaffl,M.W.法检测奶牛瘤胃主要纤维降解菌有很好的相关性(R=0.96～0.99,P<0.05);三种方法比较结果显示,SCF法省略了制作标准曲线等一系列试验操作过程,具有准确率高、快速和定量效果较好等特点。

In order to select validation of a relative quantification real-time PCR for ruminal fibrolytic bacteria, 3 healthy Chinese Holstein lactating cows with permanent ruminal cannulas were used as experimental animals,3 models for relative quantification real-time PCR were compared. The result showed that there was a close relationship between the models of SCF and the Pfafll, M.W. （R=0.96-0.99, P〈0.05 ）. In our study ,the SCF method obviated the need for construction of relative standard curves and the need for a series of validation experiments that are the prerequisite for the comparative Ct method, providing a more accurate, faster, and more efficient quantification method.