1种新型冠状病毒核酸检测试剂的性能验证  

Performance verification for a detection kit of 2019 novel coronavirus nucleic acid

作　　者：张云丽 王鑫[1] 邵玲 曲波[1] 赵鸿梅[1] ZHANG Yunli;WANG Xin;SHAO Ling;QU Bo;ZHAO Hongmei(Department of Clinical Laboratory, People′s Hospital of China Medical University, Liaoning Provincial People′s Hospital, Shenyang 110016, Liaoning, China)

机构地区：[1]辽宁省人民医院中国医科大学人民医院检验医学科,沈阳110016

出　　处：《临床检验杂志》2020年第11期827-830,共4页Chinese Journal of Clinical Laboratory Science

摘　　要：目的验证一种新型冠状病毒(2019-nCoV)核酸检测试剂盒的检测性能。方法依据CNAS-GL039《分子诊断检验程序性能验证指南》,使用2019-nCoV RNA液体性能验证参考品,依次对实时荧光RT-PCR试剂的符合率、检出限、交叉反应、精密度和抗干扰能力进行验证及评价。结果实时荧光RT-PCR试剂检测2019-nCoV RNA的符合率为100%,检出限为125 copies/mL。该试剂检测人冠状病毒HCoV-OC43、人冠状病毒HCoV-HKU1、人冠状病毒HCoV-229E、人冠状病毒HCoV-NL63、SARS冠状病毒、中东呼吸综合征(MERS)冠状病毒、甲型流感病毒、乙型流感病毒、呼吸道合胞病毒、人副流感病毒、腺病毒、肠道病毒、肺炎支原体、EB病毒、人巨细胞病毒和结核分枝杆菌无交叉反应;2.0×10^3 copies/mL和2.0×105 copies/mL的精密度参考品N基因检测的批内变异系数(CV)为0.70%及1.36%,批间CV为1.17%及1.72%;ORF1ab基因的批内CV为0.48%及0.52%,批间CV为1.36%及2.39%;加入内源性干扰物血红蛋白及清蛋白、外源性干扰物利巴韦林及阿奇霉素对2019-nCoV RNA的检测结果无影响。结论该试剂检测2019-nCoV RNA的符合率、检出限、交叉反应、精密度和抗干扰能力均符合临床分子生物学扩增检验的要求,可为临床提供可靠依据。Objective To verify the detection performance of a detection kit for 2019 novel coronavirus(2019-nCoV)nucleic acid.Methods According to the CNAS-GL039“Guidance on the Performance Verification for Molecular Diagnostic Procedures”,the coincidence rate,detection limit,cross reaction,precision and anti-interference ability of the real-time fluorescent reverse transcription polymerase chain reaction(RT-PCR)detection reagents were verified and evaluated using the reference materials of 2019-nCoV RNA.Results The coincidence rate and detection limit of the real-time fluorescent RT-PCR reagent kit for the detection of 2019-nCoV RNA were 100%and 125 copies/mL,respectively.There was no cross reaction between 2019-nCoV and human coronavirus HCoV-OC43,HCoV-HKU1,HCoV-229E,HCoV-NL63,SARS coronavirus,Middle East respiratory syndrome(MERS)virus,influenza A virus,influenza B virus,respiratory syncytial virus,human parainfluenza virus,adenovirus,enterovirus,Mycoplasma pneumonia,Epstein-Barr virus,human cytomegalovirus and Mycobacterium tuberculosis.The intra-assay coefficients of variation(CV)of the N gene for 2.0×10^3 copies/mL and 2.0×105 copies/mL of reference materials were 0.70%and 1.36%,respectively,and the inter-assay CVs were 1.17%and 1.72%,respectively.The intra-assay CVs of the ORF1ab gene were 0.48%and 0.52%,respectively,and inter-assay CVs were 1.36%and 2.39%,respectively.Endogenous interferers such as hemoglobin and albumin and exogenous interferers such as ribavirin and azithromycin had no effects on the detection of 2019-nCoV RNA.Conclusion The coincidence rate,detection limit,cross reaction,precision and anti-interference ability of the real-time fluorescent RT-PCR reagent kit for the detection of 2019-nCoV RNA meet the requirements of CNAS-GL039,which can provide a reliable basis for clinical diagnosis.

关 键 词：新型冠状病毒 核酸检测 性能验证 实时荧光聚合酶链反应 

分 类 号：R446.5[医药卫生—诊断学]