一种国产新型冠状病毒核酸检测试剂盒的性能验证

Performance Evaluation of A Domestic New Coronavirus Detection Kit

目的评估一种国产新型冠状病毒(SARS-CoV-2)核酸检测试剂盒是否满足临床SARS-CoV-2核酸检测要求。方法采用15例SARS-CoV-2核酸阳性和15例SARS-CoV-2核酸阴性的临床样本,1支SARS-CoV-2 RNA干粉标准品,5支阳性定值参考品和5支阴性定值参考品,对该试剂盒检测SARS-CoV-2阴性和阳性符合率、精密度、检出限和交叉反应进行验证。结果20例阴性和20例阳性样本的N基因和ORF1ab基因阴阳符合率均为100%。2例阳性样本ORF1ab基因重复性CV为0.41%~0.56%,优于N基因重复性,N基因和ORF1ab基因重复性、实验室总不精密度CV均<5%。检出限重复20次的检出率为100%,SARS-CoV-2与常见呼吸道病原体、与SARS-CoV-2同种属病毒(人副流感病毒1型、鼻病毒、MERS,SARS,甲型流感病毒、乙型流感病毒、冠状病毒HKU1,NL63,229E和OC43)之间不产生交叉反应。结论该试剂盒阴阳性符合率一致,重复性好。SARS-CoV-2与常见呼吸道病原体,与SARS-CoV-2同种属病毒不会产生交叉反应,适用于临床实验室和检测机构进行SARS-CoV-2核酸筛查。

Objective To evaluate whether a domestic novel coronavirus(SARS-CoV-2)nucleic acid detection kit meets the requirements of clinical new coronavirus nucleic acid detection.Methods Fifteen positive and negative clinical samples,one SARS-CoV-2 RNA dry powder standard,and five positive and negative reference samples were collected to verify the negative and positive coincidence rate,precision,detection limit,and cross-reactivity for SARS-CoV-2.Results The negative and positive coincidence rate of both N gene and ORF1ab gene were 100%(20 negative and positive samples).CV of ORF1ab gene repeatability was 0.41%~0.56%and better than that of N gene.The CV of repeatability and total laboratory imprecision of both N gene and ORF1ab gene were all less than 5%.The detection rate was 100%with 20 repeats.Common respiratory pathogens and viruses of the same genus as the new coronavirus(human parainfluenza virus type 1,rhinovirus,MERS,SARS,influenza a virus,influenza b virus,coronavirus HKU1,NL63,229E,OC43)were no cross-reaction with the SARS-CoV-2.Conclusion The kit had the same negative and positive coincidence rate,and good reproducibility.SARS-CoV-2 did not have cross-reaction with common respiratory pathogens and viruses of the same species as the SARS-CoV-2.It is suitable for clinical laboratory and testing institutions for SARS-CoV-2 nucleic acid screening.