非洲猪瘟病毒实时荧光定量PCR检测方法的建立

Establishment of qPCR detection method for African swine fever virus

为了实现非洲猪瘟病毒(African swine fever virus,ASFV)的快速检测,本研究建立了检测ASFV K196R基因的q PCR方法。按照K196R基因保守序列,设计1对特异性引物,建立并优化qPCR反应体系和反应条件,构建标准曲线,测定该检测方法的特异性、灵敏性和可重复性。结果显示,本研究建立的检测方法不与其他病毒的核酸样品产生交叉反应,检测灵敏度达10 copies/μL。变异系数小于2.5%,构建的标准曲线线性关系良好。上述结果表明,该检测方法特异性强、灵敏度高、重复性好,能够为ASF疫情防控提供技术支撑。

In order to realize the rapid detection of African swine fever virus(ASFV),a real-time PCR method was established for the detection of ASFV K196R gene.According to the conserved sequence of ASFV K196R gene,a pair of specific primers was designed to establish q PCR reaction system,with optimizing the reaction conditions and constructing the standard curve to evaluate the specificity,sensitivity and repeatability of the detection method.The results showed that the detection method did not cross reaction with other viral nucleic acid samples,and the detection sensitivity was 10 copies/μL.The coefficient of variation is less than 2.5%,and the standard curve has a good linear relationship.The detection method has the advantages of strong specificity,high sensitivity and good repeatability.It can be used for the detection of ASFV infection,then provide technical support for the prevention and control of ASF.