4种非洲猪瘟核酸检测试剂盒性能比较分析

Comparison and analysis of the detection performance of four ASFV nucleic acid detection kits

目的:为猪场实验室提供科学的非洲猪瘟病毒核酸诊断试剂盒选择方案。方法:选择4种非洲猪瘟核酸检测试剂盒,通过对比梯度稀释后标准物质的线性扩增、对低含量时(20 copies/μL)的敏感性、对以纯水为样本时假阳性检出率。结果:线性扩增试验中,A试剂盒对20 copies/μL浓度的标准品检测率为100%(3/3),其他试剂盒对该浓度下的标准品,均未达到100%;低含量敏感性试验中,A试剂盒阳性检出率为91.67%(44/48),其他厂家试剂盒的检出率分别为85.42%(41/48)、79.17%(38/48)、58.33%(28/48);假阳性试验中,A和B未出现阳性反应,而C、D试剂盒分别出现2、1个非特异性反应。结论:经过性能比较,A试剂盒更符合猪场实验室对于高敏感性和低假阳性的要求。

Objective:To provide a scientific method for nucleic acid detection of African swine fever virus(ASFV)in pig farm diagnostic laboratory.Methods:To evaluate the four selected ASFV nucleic acid detection kits,the linear amplification assay of reference substance with serial dilution,the sensitivity assay of clinical samples in low concentration(20 copies/μL)and the detection rate of false positive in pure water samples were conducted by employing the four ASFV nucleic acid detection kits.Results:In the linear amplifi cation test,the detection rate of kit A was 100%(3/3)at 20 copies/μL,while the detection rates of other kits were less than 100%;In the sensitivity test,the positive detection rate of kit A was 91.67%(44/48),and the positive rates of other kits were 85.42%(41/48),79.17%(38/48)and 58.33%(28/48),respectively;In the detection of pure water samples,kits A and B showed no positive reaction,while kits C and D showed 2 and 1 reaction with nonspecifi c amplifi cation,respectively.Conclusion:In comparison,the kit A with high sensitivity and low false positive rate is more suitable for application in laboratory of pig farms.