摘要
目的:通过分析13个SARS-CoV-2核酸检测体系临床阳性样本病毒载量与Ct值相关性及偏倚,探讨不同检测体系的结果可比性、检出限及试剂选择策略。方法:选择13个SARS-CoV-2核酸检测体系,以中国计量科学研究院标准物质倍比稀释作为标准曲线与已经过数字PCR方法赋值的临床阳性样本同时检测,采用SPSS22.0统计分析,Graph-PadPrism7.0及Medcale15.0软件绘图,统计分析临床阳性样本Ct值与病毒载量相关性、与赋值浓度的偏倚及13个检测体系的实际检出限。结果:13种检测体系中均显示病毒载量与Ct值之间相关性强,ORF1ab、N、E基因的线性回归系数分别为0.8400~0.9921、0.8674~0.9668、0.8717~0.9768;配套体系同种扩增试剂磁珠提取法优于核酸释放剂免提取法;非配套体系同种试剂不同检测体系线性回归相关系数存在较大差异。用标准曲线计算临床阳性样本ORF1ab、N、E基因结果存在差异;临床阳性样本中N基因的检出浓度高于ORF1ab的检出浓度;与数字PCR结果赋值存在较大偏倚,无法通过偏倚<7.5%,通过率≥80%的标准。利用标准曲线计算13种检测体系ORF1ab、N、E基因的检出限,与各试剂宣称的检出限之间存在差异,各体系的检测结果可比性差。结论:SARS-CoV-2核酸检测试剂检测基因的线性相关性配套体系优于非配套体系,实验室应尽可能选用配套检测体系,否则应摸索最优的检测体系。临床阳性样本的结果与赋值结果存在较大偏倚,各试剂检出限宣称值与真实值之间存在较大差异,试剂间可比性差。在暂无法实现SARS-CoV-2检测的标准化的情况下,建议实验室通过标准物质绘制标准曲线确定检测体系实际检出限的方式判断试剂真实检出限,指导初复检试剂的确定。
Objective:To analyze the correlation and bias between the viral load and Ct value of 13 SARS-CoV-2 nucleic acid detection systems,and to explore the comparability of results,detection limit of different detection systems and reagent selection strategy.Methods:Thirteen SARS-CoV-2 nucleic acid detection systems were selected,and the clinical positive samples assigned by digital PCR were detected at the same time with the standard curve of the multiple dilution of the reference material of Chinese National Institute of Metrology.SPSS 22.0 was used for statistical analysis.Graph-Pad Prism 7.0 and Medcale 15.0 software were used for mapping.The correlation between Ct value and viral load,the bias between CT value and assigned concentration,and the actual detection limit of 13 detection systems were statistically analyzed.Results:There was a strong correlation between viral load and Ct values in all 13 detection systems.The linear regression coefficients of ORF1ab,N and E genes were 0.8400-0.9921,0.8674-0.9668 and 0.8717-0.9768,respectively.The magnetic bead extraction method of matching system identical amplification reagent is better than the free extraction method of nucleic acid releasing agent.The linear regression correlation coefficients of different detection systems of the same reagent in the non-matching system were significantly different.The results of ORF1ab,N and E genes were different in clinical positive samples calculated by standard curve.The detection concentration of N gene in clinical positive samples tended to be higher than that of ORF1ab gene.There was a large bias between the values assigned to the digital PCR results,which could not pass the criteria of bias<7.5%and pass rate≥80%.The detection limits of ORF1ab,N and E genes of the 13 detection systems were calculated by standard curves.There were differences between the detection limits of ORF1Ab,N and E genes of the 13 detection systems and the declared detection limits of each reagent,and the detection results of each system were not compa
作者
曾艳芬
曹鹏驹
李瑶
陈喜军
吴泉明
陈发林
ZENG Yanfen;CAO Pengju;LI Yao(Fujian Provincial Center for Clinical Laboratory,Fujian Provincial hospital,Fujian Fuzhou 350001)
出处
《医学检验与临床》
2023年第2期1-7,共7页
Medical Laboratory Science and Clinics
基金
福建省卫生健康委新冠疫情防控科研攻关项目,项目编号:20212D02001。
关键词
新型冠状病毒
核酸检测
检测体系
相关性
偏倚
SARS-CoV-2
Nucleic acid detection
Detection system
Correlation
Bias
