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。
ObjectiveTo 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.
MethodsA 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.
ResultsThe 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).
ConclusionThis reagent of 2019-nCoV antibodies detection (colloidal gold technique) fulfills the requirement for clinical application with high specificity and sensitivity, which can be served as a supplementary detection method for 2019-nCoV nucleic acid detection by RT-PCR.
李晖,李咏茵,张志高,等. 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[J]. 中华传染病杂志,2020,38(03)：139-144.
DOI:10.3760/cma.j.issn.1000-6680.2020.03.003版权归中华医学会所有。
未经授权，不得转载、摘编本刊文章，不得使用本刊的版式设计。
除非特别声明，本刊刊出的所有文章不代表中华医学会和本刊编委会的观点。
注：2019-nCoV为2019新型冠状病毒
注：M为DNA标志物；泳道1为核蛋白基因扩增产物
注：M为蛋白质标志物；泳道1为诱导前样品；泳道2为诱导后样品
注：SDS-PAGE为十二烷基硫酸钠-聚丙烯酰胺凝胶电泳。M为蛋白质标志物；泳道1和2均为纯化后样品；泳道3为纯化样品蛋白质印迹法鉴定结果
注：2019-nCoV为2019新型冠状病毒；Ig为免疫球蛋白
| 组别 | 例数 | 2019-nCoV核酸阳性率 | 2019-nCoV IgM阳性率 | 2019-nCoV IgG阳性率 |
|---|---|---|---|---|
| 核酸阳性的确诊患者 | 89 | 89(100) | 70 (78.7) | 65 (73.0) |
| 核酸阴性的疑似患者 | 189 | ND | 113 (59.8) | 100 (52.9) |
| 对照体检者 | 273 | NA | 5 (1.8) | 2 (0.7) |
注：2019-nCoV为2019新型冠状病毒；Ig为免疫球蛋白。ND为低于检测下限；NA为不适用
李晖、李咏茵：数据整理、统计学分析、论文撰写；张志高、林冠峰、翟向明：蛋白质纯化和试剂盒生成；卢振、王奕、安泰学、胡秀梅：实验操作；赖钦涛、易璇、刘智泓、孙剑、郭亚兵、陆家韬：收集分析患者临床数据；张小勇：实验操作、研究指导、论文修改；吴英松：蛋白纯化和试剂盒生成、研究指导、论文修改；侯金林：研究指导、论文修改
文档即答
智问智答
机器翻译
你好，我可以帮助您更好的了解本文，请向我提问您关注的问题。
