评价水痘-带状疱疹病毒抗体的新型酶联免疫吸附方法  

New Enzyme-linked Immunosorbent Assay for Detection of Antibody to Varicella-zosterVirus

作　　者：权娅茹[1] 陈震[1] 邱平[1] 任晋楷 万文妍 李长贵[1] 陈国庆[1] Quan Yaru;Chen Zhen;Qiu Ping;Ren Jinkai;Wan Wenyan;Li Changgui;Chen Guoqing(National Institutes for Food and Drug Control,Beijing 102629,China;Beijing Wantai Biological Pharmaceutical Co.,Ltd.,Beijing 102206,China)

机构地区：[1]中国食品药品检定研究院,北京102629 [2]北京万泰生物药业股份有限公司,北京102206

出　　处：《中国药事》2024年第8期899-905,共7页Chinese Pharmaceutical Affairs

基　　金：国家科技重大专项-国家重大新药创制(编号2018ZX09101001-002)。

摘　　要：目的:水痘-带状疱疹病毒(VZV)糖蛋白已被证明是高度准确的VZV特异性免疫指标。本研究以重组蛋白作为包被抗原建立了酶联免疫吸附方法(ELISA),用于评价水痘疫苗接种后的血清抗体水平。方法:筛选ELISA包被抗原,以重组蛋白gE和gB作为包被抗原,W1044国际参考品作为标准品,初步建立VZV抗体的ELISA法并对其进行验证。将建立的ELISA法与膜抗原荧光抗体(FAMA)法进行比对。结果:以gE和gB作为包被抗原,成功建立了ELISA方法,采用线性拟合,标准曲线在抗体浓度0.25~4 mIU·mL^(-1)范围内具有良好的线性关系,R^(2)>0.990,回收率为85%~120%,定量限为0.25mIU·mL^(-1),精密度CV<15%,特异性良好。与FAMA法相比,ELISA法的灵敏度为89%,特异性为93%,总符合率为90%。结论:本课题成功建立了VZV抗体ELISA检测方法,可作为一种灵敏、特异的水痘疫苗临床血清评价方法。Objective:Varicella-zoster Virus(VZV)glycoproteins are the antigens that have been demonstrated to be a highly accurate indicator of VzV-specific immunity.This study established an enzyme-linked immunosorbent assay(ELISA)using recombinant glycoproteins as the coated antigens for evaluation of the serum antibody level after varicella vaccination.Methods:The ELISA coated antigen was screened,and then the recombinant proteins gE and gB were used as the coated antigens,and the W1044 international reference substance was used as the standard,and the ELISA method of VZV antibody was preliminarily established and verified.The established ELISA method was compared with the fluorescent antibody to membrane antigen(FAMA)method.Results:Using gE and gB as coated antigens,ELISA method was successfully established.Using linear fitting,the standard curve showed a good linear relationship within the antibody concentration range of 0.25-4 mIU·mL^(-1),R^(2)>0.990,with 85%-120% recovery rate,0.25 mIU·mL^(-1) limit of quantification,and less than 15% of precision CV.Compared with the FAMA method,the ELISA showed a sensitivity of 89%,specificity of 93%and overall conformity rate of 90%.Conclusion:This study successfully established a VZV antibody ELISA detection method,which can be used as a sensitive and specific quantitative method to evaluate clinical serum antibody level after varicella vaccination.

关 键 词：水痘 糖蛋白 酶联免疫吸附方法 血清学评价 

分 类 号：R917[医药卫生—药物分析学]