摘要
目的:分析3种胶体金免疫层析试剂盒检测新型冠状病毒(2019-nCoV)免疫球蛋白M(IgM)和免疫球蛋白G(IgG)抗体的特异性,探讨假阳性的干扰因素和避免方法。方法:回顾性收集2020年3月6日至4月20日北京大学第三医院门诊或住院患者的血清:(1)健康对照组120例;(2)病原感染组139例:乙型肝炎大三阳和小三阳42例、丙型肝炎抗体阳性27例、梅毒抗体阳性11例、巨细胞病毒抗体阳性12例、风疹病毒抗体阳性12例、EB病毒抗体阳性8例、甲型和乙型流感病毒患者血清各15例、大肠埃希菌血流感染患者血清12例;(3)内源性干扰组242例:血清总IgM升高20例、IgG升高20例、总补体升高15例、甲胎蛋白升高9例、类风湿因子正常组(≤20 IU/ml)84例和升高组(>20 IU/ml)94例;(4)孕妇和肿瘤患者组126例:包括孕妇、实体器官恶性肿瘤和血液系统恶性肿瘤各42例。采用逆转录聚合酶链反应(RT-PCR)检测患者咽拭子2019-nCoV核酸,阴性样本且患者排除2019-nCoV感染的纳入抗体假阳性研究。用A、B、C 3种胶体金免疫层析试剂盒检测上述血清2019-nCoV抗体,采用Pearson卡方检验对结果进行统计学分析。结果:A试剂检测健康对照血清2019-nCoV IgM和IgG的假阳性率为0和0.83%(1/120),B试剂假阳性率为4.17%(5/120),C试剂无假阳性。3种试剂检测大肠埃希菌血流感染患者血清假阳性率均为1/12,B试剂在丙肝抗体阳性血清的假阳性率为7.41%(2/27)。B试剂在血清类风湿因子正常组和升高组假阳性率分别为9.52%(8/84)和54.26%(51/94)(χ^2=40.05,P<0.001);A试剂和C试剂在类风湿因子正常组均无阳性,升高组各有3例IgM/IgG抗体阳性。A、B、C试剂在总体人群中的假阳性率分别为0.55%(3/548)、4.20%(23/548)、0.73%(4/548)。结论:不同试剂盒的特异性差别较大,高类风湿因子是导致B试剂盒假阳性的主要因素,使用高纯度的抗体和适当的阻断剂有助于提高其检测的特异度。
Objective To investigate the specificity of three gold immunochromatography assay(GICA)kits for IgM and IgG antibodies against 2019 novel coronavirus(2019-nCoV),and explore the interference factors causing false-positive results and methods for eliminating interference.Methods The serum samples were retrospectively collected from the patients presented to the Peking University Third Hospital from March 6,2020 to April 20,2020.Three commercial GICA kits were used to detect SARS-CoV-2 antibodies in serum samples from the following 4 groups.①Healthy control group:120 serum samples.②Special pathogen group:139 samples,including 42 Hepatitis B virus(HBV)antibody positive sera,27 Hepatitis C virus(HCV)antibody positive sera,11 Treponema Pallidum(TP)antibody positive sera,12 Cytomegalovirus(CMV)IgM positive sera,12 Rubella virus(RV)IgM positive sera,8 Epstein-Barr virus(EBV)IgM positive sera,12 sera with Escherichia coli confirmed by blood cultures,15 sera from influenza A virus infected patients and 15 influenza B virus infected patients.③Endogenous interference group:242 samples,including 20 sera with elevated total IgM,20 sera with elevated total IgG,15 sera with elevated total complement,9 sera with elevated alpha fetoprotein(AFP),84 sera with normal rheumatoid factor(RF)level and 94 sera with elevated RF level.④Pregnant women and patients with malignant tumor:126 samples.Reverse transcription polymerase chain reaction(RT-PCR)was used to detect 2019-nCoV viral nucleic acid in the throat swabs.Statistical analysis was performed by SPSS,version 19.0.Results The false-positive rates of 2019-nCoV IgM and IgG in healthy controls tested by A kit were 0 and 0.83%(1/120),4.17%(5/120)by B kit,and 0 by C kit.One in 12 serum samples collected from patients with E.coli bactereia was positive for 2019-nCoV antibodies tested by three kits.The false-positive rate of 2019-nCoV antibodies in HCV antibody positive sera was 7.41%(2/27)by B kit.The false-positive rates of 2019-nCoV antibodies in sera with normal and elevated RF level tested by B kit were 9.52%(8/84)and 54.26%(51/94)(χ^²=40.05,P<0.001),3 serum samples in elevated RF group were positive for 2019-nCoV IgM/IgG antibodies by A and C kits.The overall false-positive rates of A,B and C antibody test kits in all the subjects were 0.55%(3/548),4.2%(23/548)and 0.73%(4/548),respectively.Conclusions The specificity of GICA kits varies greatly among different manufacturers.Rheumatoid factor is the main factor leading to false-positive results of 2019-nCoV antibodies tested by B kit.The use of high-purity antibody and suitable immunoassay blocking reagent will be helpful to improve the specificity of GICA kits.
作者
肖秀美
段京京
吴思沂
杨博鑫
朱景昊
崔丽艳
Xiao Xiumei;Duan Jingjing;Wu Siyi;Yang Boxin;Zhu Jinghao;Cui Liyan(The Department of Laboratory Medicine,Peking University Third Hospital,Beijing 100191,China)
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2020年第11期1080-1085,共6页
Chinese Journal of Laboratory Medicine
关键词
免疫球蛋白M
免疫球蛋白G
胶体金
色谱法
亲和
假阳性反应
Immunoglobulin M
Immunoglobulin G
Gold Colloid
Chromatography,affinity
False positive reactions
作者简介
通信作者:崔丽艳,Email:cliyan@163.com。
