基于核蛋白的麻疹病毒IgG抗体间接ELISA检测方法的建立及应用  

Establishment and application of indirect ELISA for detection of measles virus IgG antibody based on nucleoprotein

作　　者：任虎[1] 曹蕾[2] 毛乃颖[2] 张荣波[1] 赵培蓓 李海 马若群 王慧玲[2] 张燕[2] 许文波[1,2,4] Ren Hu;Cao Lei;Mao Naiying;Zhang Rongbo;Zhao Peibei;Li Hai;Ma Ruoqun;Wang Huiling;Zhang Yan;Xu Wenbo(Medical College,Anhui University of Science and Technology,Huainan 232001,China;NHC Key Laboratory of Medical Virology and Viral Diseases,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,WHO WPRO Regional Reference Measles/Rubella Laboratory,Beijing 102206,China;Handan Center for Disease Control and Prevention,Handan 056004,China;Center for Biosafety Mega-Science,Chinese Academy of Sciences,Wuhan 430071,China)

机构地区：[1]安徽理工大学医学院,淮南232001 [2]中国疾病预防控制中心病毒病预防控制所,国家卫生健康委员会医学病毒和病毒病重点实验室,世界卫生组织西太平洋地区麻疹/风疹参比实验室,北京102206 [3]邯郸市疾病预防控制中心,056004 [4]中国科学院生物安全大科学研究中心,武汉430071

出　　处：《中华实验和临床病毒学杂志》2020年第1期61-66,共6页Chinese Journal of Experimental and Clinical Virology

基　　金：国家科技重大专项(2018ZX10732401)。

摘　　要：目的以原核系统表达的麻疹病毒核蛋白作为包被抗原,初步建立检测麻疹病毒抗体的间接ELISA方法,并将其应用于人群抗体水平的评估。方法对各项实验条件进行筛选和优化,确定间接ELISA法的操作流程,并检测了157份健康儿童和新生儿母亲血清中的抗麻疹病毒IgG抗体,与商品化麻疹病毒ELISA IgG抗体检测试剂盒进行比较。结果本研究建立的间接ELISA方法批内和批间的重复性试验变异系数均小于10%,与试剂盒检测结果相比,血清标本的总符合率为95.5%,灵敏度和特异度分别可达94.8%和98.3%。其中,两种方法检测的85份0~15岁健康儿童血清麻疹病毒IgG抗体阳性率,无论是<8月龄组或8月龄~15岁组,结果差异均无统计学意义(χ^2=0.313,P>0.05;χ^2=0.000,P>0.05),且通过本研究所建立的ELISA方法检测的麻疹病毒抗体水平表现出与试剂盒检测结果在不同年龄组一致的增减趋势,两种方法定量结果的相关系数r为0.893(P<0.001),显示出该方法具备的定量潜力。结论本研究建立的ELISA方法具有良好的稳定性,且灵敏度高、特异度强,与商品化试剂盒检测结果无明显差异,可用于麻疹病毒血清流行病学调查和疫苗免疫后人群抗体水平的评估。Objective To establish an indirect ELISA method for detection of measles virus antibody based on the nucleoprotein of measles virus expressed in prokaryotic system selected as the coating antigen.Methods The working conditions were screened and optimized to determine the operation process of indirect ELISA.The anti-measles virus IgG antibody in 157 sera of healthy children and mothers of newborn infants was detected and compared with the commercial measles virus ELISA IgG antibody detection kit.Results The result showed that the coefficient of variation of intra and inter assay repeatability of the indirect ELISA was less than 10%.Compared with the result of the commercial kit,the total concordance rate of serum samples was 95.5%,and the sensitivity and specificity were 94.8%and 98.3%,respectively.There was no significant difference between the two methods(χ^2=0.313,P>0.05;χ^2=0.000,P>0.05)in the positive rate of measles virus IgG antibody in 85 healthy children at the age of 0~15 years.The level of measles virus antibody detected by ELISA established in this study showed the same increasing and decreasing trend as that detected by the commercial kit in different age groups.The correlation coefficient r of the quantitative result was 0.893(P<0.001),which showed the quantitative potential of the method.Conclusions The ELISA method established in this study has good stability,high sensitivity and specificity,and there was no significant difference between the detection results of commercial kit and the ELISA method.It can be used for the seroepidemiological investigation of measles virus and the evaluation of antibody level of the population after vaccination.

关 键 词：麻疹病毒 核蛋白 血清 间接酶联免疫吸附试验 

分 类 号：R51[医药卫生—内科学] R44[医药卫生—临床医学]