恢复期血浆中新型冠状病毒核酸的提取试剂盒效率比较和检测方法适用性考察

Efficiency comparison of extraction kits and suitability study of detection method for 2019 novel corona virus nucleic acid in convalescent plasma

目的通过比较提取效率选择新型冠状病毒核酸提取试剂盒,并考察方法对血浆的核酸检测适用性。方法采用3种不同厂家核酸提取试剂盒对血浆中新型冠状病毒核酸进行提取,使用1种核酸检测试剂盒进行扩增和核酸检测。结果核衣壳蛋白(nucleocapcid,N)基因片段和开放阅读框1ab(open reading frame 1a/b,ORF1ab)基因片段标准曲线的线性相关系数均≥0.999。核酸提取试剂盒A对ORF1ab基因的回收率为91%,对N基因的回收率为38%,试剂盒B对ORF1ab基因的回收率为104%,对N基因的回收率为44%,试剂盒C对ORF1ab基因的回收率为0%,对N基因的回收率为12%.采用试剂盒A、B均能达到100%正确率,试剂盒C仅能达到62.5%正确率,误检均为假阴性。结论采用提取试剂盒A、B和新型冠状病毒2019-nCoV核酸检测试剂盒进行核酸检测,均能满足新型冠状病毒肺炎康复者恢复期血浆中病毒核酸的检测需求。

Objective To choose 2019 novel coronavirus(2019-nCoV)nucleic acid extraction kits by comparing their extraction efficiency and to investigate the suitability of detection method for nucleic acid in plasma sample.Methods The 2019-nCoV nucleic acid in plasma was extracted using 3 different nucleic acid extraction kits.The open reading frame 1a/b(ORF1ab)and nucelocapcid(N)genes were amplified and detected using same nucleic acid detection kit.Results The correlation coefficients of ORF1ab and N gene fragments standard curve were both≥0.999.Extraction kit A had a recovery rate of 91%for ORF1ab and 38%for N.Kit B had a recovery rate of 104%for ORF1ab and 44%for N.Kit C had a recovery rate of 0%for ORF1ab and 12%for N.The accuracies of kits A and B were both 100%,while kit C was 62.5%attributing to false negative results.Conclusion Except for kit C,both kits A and B can be used with 2019-nCoV nucleic acid detection kit for virus nucleic acid detection in COVID-19 convalescent plasma.