基于Crisper的新型冠状病毒核酸检测方法在临床应用中的评价  

An Evaluation of New Coronavirus Nucleic Acid Detection Method in Clinical Applications Based on CRISPR Method

作　　者：裴兵 孙建[1] 崔倩 戚小艳 张振江 PEI Bing;SUN Jian;CUI Qian;QI Xiaoyan;ZHANG Zhenjiang(Department of Clinical Laboratory, The Affiliated Suqian First People′s Hospital of Nanjing Medical University, Suqian 223800, China)

机构地区：[1]南京医科大学附属宿迁市第一人民医院医学检验科,江苏宿迁223800

出　　处：《标记免疫分析与临床》2021年第6期1051-1054,共4页Labeled Immunoassays and Clinical Medicine

基　　金：宿迁市科技重点实验室项目(编号:M201902)。

摘　　要：目的建立一种快速、特异性强、灵敏度高的新型冠状病毒核酸检测方法并评价其在临床工作中的应用。方法对2020年2月至5月经过RT-PCR及CT方法临床确诊的新冠患者标本,采用Crisper方法对50例正常标本及12例确诊标本进行检测,用本方法使12例确诊标本全部进行识别并检出,而用RT-PCR方法仅能检出10例,存在2例漏检的问题。结果Crisper方法特异性为100%;灵敏度为91.3%;精密度为高值批内精密度CV为0.66%,低值批内精密度CV为1.50%,均小于5%,符合试剂批内精密度要求;线性范围的相关系数r=0.9980,符合|r|≥0.980要求;可报告阳性为Ct值<37;验证方法符合要求。结论具有成本低、可多次重复检测、方法简单、检测速度快、灵敏,最低检测极限达到10-2拷贝/μL、特异,可在短时间内通过荧光信号的变化检测到是否含有目标核酸。Objective To establish and evaluate a rapid,specific and sensitive new Coronavirus nucleic acid detection method in clinical work.Methods From February to may,in 2020,50 normal samples and 12 confirmed cases were detected by CRISPR method.All 12 cases were identified and detected by this method,while only 10 cases were detected by RT-PCR,and 2 cases were missed.Results The specificity of CRISPR method was 100%,while the sensitivity was 91.3%.The precision was high-value,the intra-run precision CV was 0.66%,and the low-value intra-run precision CV was 1.50%,which was less than 5%and in line with the reagent intra-run precision requirements.The correlation coefficient of the linear range was r=0.9980,which met the requirements of|r|≥0.980.The reportable positive was Ct value<37 and the verification method met the requirements.Conclusion CRISPR method has the advantages of low cost,easy to repeat,simple and fast,high sensitivity and specificity.The lowest detection limit is 10-2 copies/μL and can detect whether the target nucleic acid is contained in a short time by the change of fluorescence signal.

关 键 词：Crisper RT-PCR方法 新型冠状病毒 特异性强 灵敏度 

分 类 号：R373.9[医药卫生—病原生物学]