摘 要：为建立一种特异性强、灵敏度高、重复性好的非洲猪瘟病毒微滴数字PCR检测方法，本研究根据非洲猪瘟病毒核酸的2段保守序列（VP72和K205基因）设计合成了3对引物和探针，摸索其反应条件，优化方法，比较方法的线性、特异性、灵敏性和重复性。结果发现，这3对引物和探针的非洲猪瘟病毒方法检出限均小于6拷贝数/反应，在10~106拷贝数/反应之间均呈良好的线性关系，定量范围内的检测变异系数均低于15%，且不与猪常见的2种病毒发生交叉反应，适用于猪肉及其制品中非洲猪瘟病毒的检测，有利于在源头、食品加工、食品销售等各个环节加强对生鲜猪肉及各类猪肉制品中非洲猪瘟病毒的检测，切实降低潜在风险。

Abstract: In order to establish African swine fever virus droplet digital PCR detection method with high specificity, high sensitivity and good repeatability, three pairs of primers and probes were synthesized based on two conserved sequences of African swine fever virus nucleic acid, VP72 and K205 genes. In this study, the method was optimized and the linearity, specificity, sensitivity and repeatability were compared. It was found that the detection limits of the three pairs of primers and probes were all less than 6 copies / reaction, and there was a good linear relationship between 10~106 copies / reaction. The coefficients of variation within the quantitative range were all less than 15%. Moreover, synthesized three pairs of primers and probes did not cross-react with the two common viruses of pigs. This method is suitable for the detection of African swine fever virus in pork and its products. It is beneficial to detect African swine fever virus in fresh pork and various pork products at various area such as source, food processing, and food sales, and effectively reduce potential risks.