猪细小病毒SYBR Green Ⅰ模式实时定量PCR检测方法的建立

Development of SYBR Green Ⅰ Real-time PCR Method for Detection of Porcine parvovirus

根据GenBank猪细小病毒(PPV)的NS1基因序列,设计一对特异性引物,采用SYBR Green Ⅰ随机结合渗入法,建立实时定量PCR检测方法,构建了检测PPV的标准DNA模版,循环阈值(Ct)与标准质粒DNA模板在7.8×101～7.8×108拷贝/μl浓度范围内呈良好的线性关系,相关系数为0.997。该方法用于猪细小病毒的检测具有很高的特异性,其敏感性与常规PCR相比可以提高100倍,可以用于猪细小病毒的快速检测。

Basd on converse regions of PPV NS1 genome sequences in GenBank, one pair of specific primer was designed. A real time quantitative PCR method is established to detect PPV standard DNA template, by using SYBR Green I random combination. The cycle threshold and standard DNA template are between 7.810^1 -7.8× 10^8 copy/ μ 1 with good linearity and, the correlation coefficient is 0.997. This method is high specific in detecting PPV, the sensitivity is 100 times than general PCR . It can be used in rapidty detection of in PPV.