2019新型冠状病毒抗体胶体金检测方法的建立与临床性能评价  

Establishment and clinical performance evaluation of 2019 novel coronavirus antibody colloidal gold detection method

作　　者：李晖[1] 李咏茵[1] 张志高 卢振 王奕 林冠峰 安泰学 胡秀梅[5] 赖钦涛[1] 易璇 刘智泓[1] 翟向明 孙剑[1] 郭亚兵[1] 陆家韬 张小勇[1] 吴英松 侯金林[1] Li Hui;Li Yongyin;Zhang Zhigao;Lu Zhen;Wang Yi;Lin Guanfeng;An Taixue;Hu Xiumei;Lai Qintao;Yi Xuan;Liu Zhihong;Zhai Xiangming;Sun Jian;Guo Yabing;Lu Jiatao;Zhang Xiaoyong;Wu Yingsong;Hou Jinlin(Department of Infectious Diseases,Nanfang Hospital,Southern Medical University,State Key Laboratory of Organ Failure Research,Guangdong Provincial Key Laboratory of Viral Hepatitis Research,Guangzhou 510515,China;Guangzhou DARUI Biotechnology Co.,Ltd,Guangzhou 510530,China;Department of Laboratory Medicine,the People′s Hospital of Honghu,Honghu City,Hubei Province 434000,China;School of Laboratory Medicine and Biotechnology,Southern Medical University,Guangzhou 510515,China;Department of Laboratory Medicine,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China;Wuhan Hankou Hospital,Wuhan 430010,China)

机构地区：[1]南方医科大学南方医院感染内科,器官衰竭防治国家重点实验室,广东省病毒性肝炎研究重点实验室,广州510515 [2]广州市达瑞生物技术股份有限公司,510530 [3]洪湖市人民医院医学检验科,湖北省434000 [4]南方医科大学检验与生物技术学院,广州510515 [5]南方医科大学南方医院检验医学科,广州510515 [6]武汉市汉口医院,430010

出　　处：《中华传染病杂志》2020年第3期139-144,共6页Chinese Journal of Infectious Diseases

摘　　要：目的:建立2019新型冠状病毒(2019 novel coronavirus,2019-nCoV)免疫球蛋白(immunoglobulin,Ig)M和IgG胶体金快速检测方法,并对检测试剂进行临床性能评价。方法:收集2020年2月12日至20日分别在武汉市汉口医院和洪湖市人民医院就诊的共278例患者,根据诊断标准分成2019-nCoV核酸阳性的确诊患者89例和2019-nCoV核酸阴性的疑似患者189例。选取2015年至2018年南方医科大学南方医院的273名体检者作为对照。采集患者血清样本。采用原核表达载体获得2019-nCoV病毒重组核蛋白,利用重组核蛋白分别建立间接法IgM和IgG胶体金检测方法,以核酸反转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)检测方法作为对照,对3组人群的血清标本进行2019-nCoV IgM和IgG检测,分析胶体金检测法的特异度和灵敏度。结果:采用胶体金试剂检测核酸阳性的确诊患者的2019-nCoV IgM和IgG阳性检出率分别为78.7%(70/89)和73.0%(65/89);体检者的2019-nCoV IgM和IgG阳性检出率分别为1.8%(5/273)和0.7%(2/273)。2019-nCoV IgM检测胶体金试剂的灵敏度和特异度分别为78.7%和98.2%,2019-nCoV IgG检测试剂的灵敏度和特异度分别为73.0%和99.3%,联合IgM和IgG检测的灵敏度和特异度分别为87.6%和98.2%。核酸阴性的疑似患者的2019-nCoV IgM和IgG阳性检出率分别为59.8%(113/189)和52.9%(100/189),IgM和IgG联合检测阳性检出率为66.1%(125/189)。结论:2019-nCoV抗体检测试剂(胶体金法)的特异度和灵敏度高,可作为已有的2019-nCoV核酸RT-PCR检测法的补充手段。Objective To establish a colloidal gold technique assay for the rapid detection of immunoglobulin(Ig)M and IgG antibodies against 2019 novel coronavirus(2019-nCoV)and to evaluate its clinical performance.Methods A total of 278 patients who were respectively treated at Wuhan Hankou Hospital and the People′s Hospital of Honghu from February 12,2020 to February 20,2020 were collected.According to the diagnostic criteria,89 patients were confirmed with positive 2019-nCoV nucleic acid,and 189 were 2019-nCoV nucleic acid-negative suspected patients.A total of 273 medical examiners from Nanfang Hospital,Southern Medical University from 2015 to 2018 were selected as controls.The serum samples of patients were collected.2019-nCoV nucleic proteins were obtained from prokaryotic expression vectors.Indirect IgM and IgG colloidal gold techniques were established by using recombinant nuclear protein.2019-nCoV nucleic acid detection by reverse transcription-polymerase chain reaction(RT-PCR)was used as control.Serum specimens were tested for 2019-nCoV IgM and IgG.The specificity and sensitivity of colloidal gold assay were analyzed.Results The positive rates of IgM and IgG with the colloidal gold detection in confirmed patients with positive 2019-nCoV nucleic acid were 78.7%(70/89)and 73.0%(65/89),respectively.The positive rates of IgM and IgG in medical examiners were 1.8%(5/273)and 0.7%(2/273),respectively.The sensitivity and specificity of IgM detection reagents were 78.7%and 98.2%,respectively,those of IgG detection reagents were 73.0%and 99.3%,respectively,and those of IgM combined with IgG detection were 87.6%and 98.2%,respectively.For suspected patients with negative 2019-nCoV nucleic acid,the positive rates of IgM and IgG were 59.8%(113/189)and 52.9%(100/189),respectively,and the positive rate of IgM combined with IgG detection was 66.1%(125/189).Conclusion This reagent of 2019-nCoV antibodies detection(colloidal gold technique)fulfills the requirement for clinical application with high specificity and sensitivity,whic

关 键 词：冠状病毒感染 2019新型冠状病毒 胶体金 抗体 临床性能 

分 类 号：R512.99[医药卫生—内科学]