柯萨奇病毒A组16型中和抗体检测方法的实验室评价  

Laboratory evaluation on test method for neutralizing antibody against coxsacievirus A16

作　　者：毛群颖[1] 邵杰[1] 罗震 高帆[1] 安文琪 郝春生 高强[4] 杨二霞 韩金乐 李振平 王一平 梁争论[1] 徐苗[1] 

机构地区：[1]中国食品药品检定研究院,北京100050 [2]华兰生物工程股份有限公司研发部,河南新乡453003 [3]北京微谷生物医药有限公司,北京100024 [4]北京科兴生物制品有限公司研发部,北京100085 [5]中国医学科学院医学生物学研究所免疫室,云南昆明650118 [6]北京万泰生物药业股份有限公司,北京102206 [7]北京智飞绿竹生物制药有限公司,北京100176

出　　处：《中国生物制品学杂志》2014年第6期829-834,842,共7页Chinese Journal of Biologicals

基　　金：国家高技术研究发展计划"863计划"资助项目(2012AA02A402)

摘　　要：目的评价柯萨奇病毒A组16型(coxsackievirus A16,CA16)中和抗体检测方法于不同实验室内和实验室间的可重复性。方法参照WHO脊髓灰质炎病毒中和抗体检测方法,建立CA16中和抗体检测方法和实验标准;收集22份血清样品,对CA16 A基因型毒株G10及B基因型毒株W190、33、203、P11和CS02毒株独立进行CA16中和抗体有效检测实验,评价所建立的检测方法于A^G实验室内和实验室间的可重复性;分别于A、B、C、D和E实验室比较G10毒株与W190、33、203、P11和CS02毒株对CA16中和抗体检测结果的影响。结果在各实验室内,有17/22份血清样品的变异系数(variable coefficient,CV)在0~11.4%之间,表明该检测方法在同一实验室内具有较好的可重复性;在实验室间,除3份低效价血清和1份效价过高血清外,其余血清样品的几何平均滴度(geometric mean titer,GMT)最大差异倍数均在3.0倍以内,实验室间检测差异在可接受范围内;W190、33、203、P11和CS02毒株与G10毒株检测结果趋势相近。在同一实验室内,W190和33毒株与G10毒株的检测结果差异无统计学意义;203、P11和CS02毒株的检测结果均低于G10毒株,差异有统计学意义,表明B基因型毒株与A基因型毒株生物学活性存在一定差异。结论以G10毒株检测CA16中和抗体的方法在各协作实验室可得到良好应用,有利于CA16中和抗体检测的标准化,但适用性需进一步评估和验证。Objective To evaluate the intra- and inter-reproducibility of test method for neutralizing antibody against coxsaeievirus A16 （CA16） in various laboratories. Methods The test method and experimental standard for neutralizing antibody against CA16 were developed according to the WHO requirements for test method for neutralizing antibody against poliovirus. Twenty-two serum samples were collected and tested for neutralizing antibody against CA16 in strain G10 of genotype A and strains W190, 33, 203, Pll and CS02 of genotype B separately to evaluate the intra- and inter- reproducibility of the method in laboratories A - G. The influences of these strains on test result of neutralizing antibody against CA16 were compared in laboratories A -E. Results The intra-coefficients of variation （CVs） of test results of 17 of the 22 samples were 0 - 11. 4%, indicating good intra-reproducibility of the method. However, except three samples with low titers and one with high titter, the difference of GMTs of other serum samples was within 3. 0 folds among seven laboratories, which was acceptable. The test results of strains W190, 33, 203, Pll and CS02 showed similar tendencies to that of strain G10. The test results of strains W190 and 33 in the same laboratory showed nosignificant difference with that of strain G10. However, the test results of strains 203, P11 and CS02 were significantly lower than that of G10, indicating a certain difference in biological activities of strains of genotypes A and B. Conclusion The test method for neutralizing antibody against CA16 of strain G10 were used effectively in the cooperative laboratories, which was beneficial to the standardization of CA16 neutralizing antibody. Howerer, the applicabilidy of the method needs to be further assessed and confirmed.

关 键 词：柯萨奇病毒A组16型 中和抗体 基因型 

分 类 号：R373.23[医药卫生—病原生物学] R392-33[医药卫生—基础医学]