2019新型冠状病毒核酸检测体系性能验证

Performance Verification of 2019 New Coronavirus Nucleic Acid Detection System

目的:评估2019新型冠状病毒(2019-nCoV)核酸检测体系在质量上是否能满足临床需求。方法:对本实验室采用的新冠核酸采样管、提取试剂、两种扩增试剂及相应仪器在配套使用时的阴阳性符合率、批内精密度、批间精密度及最低检测限、交叉反应等项目进行性能验证。结果:本实验室的新冠核酸检测体系阴阳性符合率100%;ORF1ab、N与E各基因批内精密度、批间精密度CV均≤5%;新冠标准物质稀释至厂家声明的最低检测限浓度500 copies/ml时,ORF1ab、N与E每个基因重复提取扩增10次,每次均出现了扩增曲线,Ct值均<37,检出率100%,利用本核酸检测体系对人类冠状病毒(HKU1、OC43、NL63和229E)、MERS冠状病毒、甲型流感病毒、乙型流感病毒、人副流感病毒Ⅰ型、腺病毒Ⅰ型、EB病毒、人巨细胞病毒、肺炎支原体及人基因组DNA进行提取扩增,结果为阴性。结论:本实验室的新冠核酸检测体系阴阳性符合率一致,重复性好,最低检测限与厂家声明一致,与能引起相同或相似临床症状的其他病原体及人基因组DNA不产生交叉反应,特异性好,能够在质量上保证临床新冠核酸检测的需要,可为其他实验室选择配套检测体系提供参考依据。

Objective:To evaluate whether the 2019 New Coronavirus(2019-nCoV)nucleic acid detection system can meet the clinical needs in terms of quality.Methods:The performance of the new coronal nucleic acid sampling tube,extraction reagent,two kinds of Amplification Reagents and corresponding instruments used in our laboratory were verified,including the coincidence rate of the positive and negative,intra batch precision,inter batch precision,minimum detection limit and cross reaction.Results:The positive and negative coincidence rate of the new detection system was 100%.Within run and inter run CV of ORF1ab,N and E genes were less than 5%.When diluted to 500 copies/ml,ORF1ab,N and E genes were repeatedly extracted and amplified for 10 times,and the amplification curve appeared every time.The Ct value was less than 37,and the detection rate was 100%.The nucleic acid detection system was used to detect human coronavirus(HKU1,OC43,NL63 and 229E),MERS coronavirus,Influenza A virus,Influenza B virus,human parainfluenza virus typeⅠ,adenovirus typeⅠ,Epstein Barr virus,human cytomegalovirus,Mycoplasma pneumoniae and human genomic DNA were extracted and amplified,and the results were negative.Conclusion:The positive and negative coincidence rate of the new coronal nucleic acid detection system in our laboratory is consistent,and the repeatability is good.The minimum detection limit is consistent with the manufacturer’s statement.There is no cross reaction with other pathogens and human genome DNA that can cause the same or similar clinical symptoms,and the specificity is good,which can ensure the needs of clinical new coronal nucleic acid detection in terms of quality,it can provide reference for other laboratories to choose the supporting test system.