新型冠状病毒灭活样品混合进行核酸检测效果分析  

Impact of pooling of inactivated SARS-CoV-2 samples on nucleic acid detection

作　　者：余波[1] 霍细香[1] 汪杨 江永忠[1] YU Bo;HUO Xixiang;WANG Yang;JIANG Yongzhong(Hubei Provincial Center for Disease Control and Prevention,Wuhan 430079,China;Hubei Institute for Drug Control,Wuhan 430075,China)

机构地区：[1]湖北省疾病预防控制中心,武汉430079 [2]湖北省药品监督检验研究院

出　　处：《公共卫生与预防医学》2020年第5期11-14,共4页Journal of Public Health and Preventive Medicine

基　　金：国家科技重大专项课题(2018ZX10734401-009)。

摘　　要：目的了解新型冠状病毒灭活样本混合检测对结果的影响,为大人群样本的新型冠状病毒核酸筛查提供科学检测的方案。方法将新型冠状病毒阳性咽拭子样本和阴性咽拭子样本于56℃30min灭活后,按照阳性样本1份、阴性样本4、9、19份的比例进行混合,用实时荧光RT-PCR技术检测原液和混合液的ORF-lab和N基因。结果20组灭活样品原液核酸检测结果全部为阳性,1∶5混合液和1∶10混合液核酸检测结果也均是阳性,1∶20混合液有1组样品ORF-lab阴性,N基因阳性。各组间核酸检测Ct值均显著相关。不同样本组组间阳性率差异无统计学意义。20组样品ORF-lab基因1∶5混合液、1∶10混合液、1∶20混合液相比原液Ct值分别平均增加1.73、2.86、4.05,N基因1∶5混合液、1∶10混合液、1∶20混合液相比原液Ct值分别平均增加1.69、2.79、3.25。结论当开展大人群样本的新型冠状病毒核酸筛查时,选择<10份灭活样品混合检测,大部分情况下不影响定性结果,但弱阳性样本结果可能会受到影响。Objective To investigate the impact of pooling of inactivated samples on testing results of nucleic acid detection of SARS-CoV-2,and to provide a scientific detection scheme for the SARS-CoV-2 nucleic acid screening of large population samples.Methods The SARS-CoV-2 positive throat swab samples and the negative throat swab samples were inactivated at 56°C for 30 minutes,and mixed according to the ratio of positive samples to negative samples at 1∶4,1∶9,and 1∶19,respectively.Real-time fluorescent RT-PCR technology was used to detect the ORF-lab and N genes in the original solution and mixed solution.Results The nucleic acid test results of the 20 groups of inactivated samples were all positive.The nucleic acid test results of the 1∶5 mixture and the 1∶10 mixture were also positive.One group of samples of the 1∶20 mixture was negative for ORF-lab and positive for the N gene.The Ct values of nucleic acid detection among all groups were significantly correlated.There was no statistically significant difference in the positive rate between different sample groups.Compared with the original solution,the Ct values of the ORF-lab gene of 1∶5 mixture,1∶10 mixture,and 1∶20 mixture samples increased by 1.73,2.86,and 4.05,respectively,while the Ct values of the N gene of 1∶5 mixture,1∶10 mixture and 1∶20 mixture samples increased by 1.69,2.79,and 3.25,respectively.Conclusion When conducting nucleic acid screening for SARS-CoV-2 in large population samples,a mixed test of less than 10 inactivated samples would not affect the qualitative results in most cases,but the results of weak positive samples may be affected.

关 键 词：新型冠状病毒 灭活 样品混合 核酸检测 

分 类 号：R183[医药卫生—流行病学]