水痘-带状疱疹病毒IgG抗体ELISA定量检测方法的优化、验证及应用  

Optimization,verification and application of ELISA method for quantitative detection of varicella-zoster virus IgG antibody

作　　者：周志军 陈克金 林连珍 李娟 王敏 陈丽梅 程爽 罗敏华[2] 曾双迎 李陶敬 胡勇 李策生 ZHOU Zhijun;CHEN Kejin;LIN Lianzhen;LI Juan;WANG Min;CHEN Limei;CHENG Shuang;LUO Minhua;ZENG Shuangying;LI Taojing;HU Yong;LI Cesheng(Sinopharm Wuhan Plasma-derived Biotherapies Co.,Ltd.,Wuhan 430207,Hubei Province,China;不详)

机构地区：[1]国药集团武汉血液制品有限公司,湖北武汉430207 [2]中国科学院武汉病毒研究所,湖北武汉430207 [3]成都蓉生药业有限责任公司,四川成都610041

出　　处：《中国生物制品学杂志》2023年第8期973-979,共7页Chinese Journal of Biologicals

基　　金：国家科技部重点专项(2020YFC0841800)。

摘　　要：目的 优化水痘-带状疱疹病毒(varicella-zoster virus,VZV)IgG抗体效价ELISA定量检测方法,进行验证后用于健康人群中高效价VZV-IgG血浆筛选。方法 选用德国维润赛润研发有限公司VZV-IgG间接ELISA试剂盒,将第1代VZV人免疫球蛋白国际标准品(NIBSC编号:W1044)稀释至2 IU/mL作为标准品,采用4参数拟合曲线建立定量ELISA法,确定其最佳线性范围;对优化后的方法进行精密度、准确度验证。用建立的方法检测国药集团武汉血液制品有限公司辖属10个浆站1 962份人血浆样品VZV-IgG抗体效价,以及部分批次人免疫球蛋白制剂。结果 标准曲线线性范围为16.25~2 000 mIU/mL,批内精密度CV为1.3%~10.6%,批间精密度CV为4.270%~7.636%,批内准确度为92.30%~111.02%,批间准确度为98.4%~104.88%;样品添加试验显示,添加样品的实测值为理论值的95.79%~111.03%。1 962份人血浆样本的阳性率为94.29%,效价大于3 000 mIU/mL占比1.02%。不同种类人免疫球蛋白制剂中VZV-IgG抗体效价均较低,且均高于静注人免疫球蛋白(pH 4)。结论 优化后的VZV-IgG定量检测方法可用于健康人群VZV-IgG筛查,自然感染的健康献浆员的VZV-IgG抗体阳性率高,但效价低,需进行疫苗免疫获得合格的高效价血浆。Objective To optimize and verify the ELISA method for quantitative detection of varicella-zoster virus(VZV)IgG antibody potency,and use it for the screening of plasma with high potency VZV-IgG in healthy donors.Methods The VZV-IgG indirect ELISA kit from Institut VirionSerion GmbH was selected,the first international standard for varicellazoster immunoglobulin(NIBSC code:W1044)was diluted to 2 IU/mL as the standard,and 4-parameter fitting curve was used to develop the quantitative ELISA method. The method was determined for the optimal linear range and verified for the precision and accuracy. VZV-IgG antibody potency of 1 962 human plasma samples and some batches of human immunoglobulin preparations from 10 plasma stations under Sinopharm Wuhan Plasma-derived Biotherapies Co.,Ltd.(SWPB)were detected by the developed method.Results The linear range of the standard curve was 16. 25 ~ 2 000 mIU/mL,the CV values of precision in intra-and inter-assays were 1. 3% ~ 10. 6% and 4. 270% ~ 7. 636%,and the accuracy in intra-and inter-assays were 92. 30% ~ 111. 02% and 98. 40% ~ 104. 88%,respectively;Sample-adding experiment showed that the measured value of the added sample was 95. 79% ~ 111. 03% of the theoretical value. The positive rate of 1 962 human plasma samples was 94. 29%,and the samples with potency greater than 3 000 mIU/mL accounted for 1. 02%. The potency of VZV-IgG antibody in different kinds of human immunoglobulin preparations was lower,while higher than that of intravenous human immunoglobulin(pH 4).Conclusion The optimized VZV-IgG quantitative detection method can be used for the screening of VZV-IgG in healthy people. The positive rate of VZV-IgG antibody in naturally infected healthy plasma donors is high,while the potency is low,thus,vaccine immunization is required to obtain qualified plasma with high potency.

关 键 词：水痘-带状疱疹病毒 IGG ELISA 阳性率 效价 

分 类 号：Q599[生物学—生物化学] R34[医药卫生—基础医学]