新型冠状病毒核酸提取方法的性能评价及临床应用  

Performance evaluation and clinical application of Severe Acute Respiratory Syndrome Coronavirus 2 nucleic acid extraction methods

作　　者：吴永彬 万颖 蒋红玲[1] 张晶[1] 辛杰 WU Yongbin;WAN Ying;JIANG Hongling;ZHANG Jing;XIN Jie(Department of Clinical Laboratory,Nanxishan Hospital of Guangxi Zhuang Autonomous Region,Guilin,Guangxi 541002,China)

机构地区：[1]广西壮族自治区南溪山医院检验科,广西桂林541002

出　　处：《检验医学与临床》2021年第6期725-728,共4页Laboratory Medicine and Clinic

摘　　要：目的评价3种新型冠状病毒(SARS-CoV-2)核酸提取方法的分析性能及临床应用价值。方法采用目前临床常用的裂解法、柱提法和磁珠法提取SARS-CoV-2核酸,通过实时荧光反转录聚合酶链反应及自建检测系统检测SARS-CoV-2核酸[开放阅读框(ORF)1ab、核衣壳蛋白(N)],评价3种核酸提取方法下的检测精密度、正确度、最低检测限等性能指标。将3种核酸提取方法应用于17份新型冠状病毒肺炎(COVID-19)确诊病例标本的检测,分析其临床应用价值。结果3种核酸提取方法下检测结果的重复性精密度和中间精密度、正确度均通过验证,可满足临床需求。采用裂解法、柱提法、磁珠法提取核酸时的最低检测限(Ct值)分别是37.42、37.87、39.61,均符合厂家声明的最低检测限。在应用于临床标本检测时,3种核酸提取方法下的检测结果两两之间均呈良好的线性关系(P<0.05),其中,采用柱提法和磁珠法提取核酸时的检测结果相关性最好(r=0.905,P<0.001)。3种核酸提取方法下的靶标(ORF1ab+/N-、ORF1ab-/N+、ORF1ab+/N+)检出率比较,差异无统计学差异(P>0.05)。结论裂解法、柱提法和磁珠法提取SARS-CoV-2核酸在自建检测系统中的分析性能可接受,均可满足临床对SARS-CoV-2的检测需求。Objective To evaluate the analytical performance and clinical application value of 3 Severe Acute Respiratory Syndrome Coronavirus 2(SARS-CoV-2)nucleic acid extraction methods.Methods The SARS-CoV-2 nucleic acid was extracted by the lysis method,column extraction method and magnetic bead method commonly used in clinical practice.SARS-CoV-2 nucleic acids[open reading frame(ORF)1ab,nucleocapsid protein(N)]were detected by real-time fluorescent reverse transcription polymerase chain reaction and self-designed detection system.Evaluated the precision,accuracy and minimum detection limit of the 3 methods.Three nucleic acid extraction methods were applied to the detection of specimens from 17 confirmed cases of Corona Virus Disease 2019(COVID-19),and their clinical application value was analyzed.Results The repeatability precision,intermediate precision and accuracy of the detection results of the 3 nucleic acid extraction methods all verified,and met the clinical needs.The minimum detection limits(Ct value)for nucleic acid extraction by lysis method,column extraction method and magnetic bead method were 37.42,37.87,39.61 respectively,which met the minimum detection limits declared by the manufacturer.When applied to clinical specimen detection,the detection results of the 3 nucleic acid extraction methods showed a good linear relationship(P<0.05).Among them,the correlation between the results of nucleic acid extraction by column extraction method and magnetic bead method was the best(r=0.905,P<0.001).There was no significant difference in the detection rate of targets(ORF1ab+/N-,ORF1ab-/N+,ORF1ab+/N+)of the 3 nucleic acid extraction methods(P>0.05).Conclusion The analytical performance of the lysis method,column extraction method and magnetic bead method to extract SARS-CoV-2 nucleic acid in the self-designed detection system is acceptable,and can meet the clinical requirements for SARS-CoV-2 detection.

关 键 词：新型冠状病毒 核酸提取 实时荧光反转录聚合酶链反应 

分 类 号：R446.5[医药卫生—诊断学]