人脑膜炎球菌参考血清 Men10的制备及其抗A、C、Y、W135群脑膜炎血清群荚膜多糖抗体IgG含量和体外杀菌试验滴度  

Preparation of a human meningococcal reference serum and standardization of IgG concentrations to capsular polysaccharides and bactericidal activities against serogroup A, C, Y and W135 strains

作　　者：赵志强[1] 王欣茹[1] 李亚南[2] 王浩[1] 刘方蕾[1] 袁菲[1] 乔瑞洁[1] 蒋井明[1] 何彦林[1] 林纪胜[3] 叶强[2] 谢贵林 

机构地区：[1]国药中生兰州生物制品研究所有限责任公司,甘肃省疫苗工程技术研究中心,730046 [2]中国食品药品检定研究院 [3]美国阿拉巴马大学伯明翰分校病理学系呼吸道病原参考实验室、WHO肺炎链球菌参考实验室 [4]中国生物技术股份有限公司,北京100029

出　　处：《中华微生物学和免疫学杂志》2014年第6期459-464,共6页Chinese Journal of Microbiology and Immunology

基　　金：国家科技支撑计划课题（2008BAI66B01）

摘　　要：目的：制备人脑膜炎球菌参考血清，对其中A、C、Y、W135血清群的抗荚膜多糖抗体IgG含量和体外杀菌试验（ serum bactericidal assay ， SBA）滴度进行定值。方法用4价A、C、Y、W135群脑膜炎荚膜多糖疫苗免疫20名健康成人，采集免疫后的血浆进行混合、去纤维蛋白原、过滤、分装、冻干，得到人脑膜炎球菌参考血清Men10。在兰州生物制品研究所有限责任公司（ LIBP ）实验室，用标准检测人血清中抗脑膜炎球菌荚膜多糖抗体IgG定量ELISA方法，以国际标准血清CDC1992为标准，对其中4个血清群抗荚膜多糖抗体IgG含量定值；以临时定值的Men10和CDC1992分别为标准检测12份美国疾病预防控制中心（ CDC）的校正血清、56份4价多糖疫苗免疫后的血清，通过比较对定值的准确性进一步验证。同时以体外杀菌试验测定针对4个血清群的SBA滴度。结果制备了4000支（0．5 ml/支）冻干人脑膜炎球菌参考血清Men10。该血清HBV、HCV、HIV、梅毒螺旋体检测均阴性，残留水分含量2．3％。以Men10为标准检测的12份CDC校正血清和56份4价多糖疫苗免疫后的血清群抗荚膜多糖抗体结果，与以CDC1992为标准检测的结果均具有良好的直线相关关系（ r＝0．99，P＜0．05）；以Men10为标准检测的CDC1992的4个血清群抗荚膜多糖抗体的新值与其原定值比较，各血清型的误差均＜10％。同时确立了Men10中各血清群的体外杀菌试验滴度。结论 LIBP成功制备了人脑膜炎球菌参考血清Men10并完成了对其中的4个血清群抗荚膜多糖抗体IgG含量的准确定值，并确定了其中各群别抗体的体外杀菌试验滴度。Objective To prepare a human meningococcal reference serum and standardize IgG concentrations to capsular polysaccharides and in vitro bactericidal activities of the reference serum against serogroup A, C, Y and W135 strains.Methods Twenty healthy adults were recruited and given one dose of immunization with tetravalent （serogroups A, C, Y and W135） meningococcal polysaccharide vaccine . Plasma samples were collected and gone through a series of process treatments including defibrination , filtration, and lyophilization to prepare the meningococcal reference serum Men 10.The IgG concentrations of Men10 to capsular polysaccharides of serogroups A , C, Y and W135 were calibrated by using an international reference serum CDC1992 as the standard in enzyme-linked immunosorbent assay （ELISA）.Provisional IgG concentrations of Men10 were intensively validated by testing a panel of 12 calibration serum samples from Centers for Disease Control and Prevention , USA （ US CDC） and a panel of 56 serum samples immunized with A, C, Y and W135 meningococcal polysaccharide vaccine from Lanzhou Institute of Biological Products Co., Ltd.（LIBP） with the assays using Men10 and CDC1992 as the standard and/or test samples, respectively.The bactericidal titers against serogroup A , C, Y and W135 strains were measured by serum bactericidal assay （SBA）.Results Four thousand vials （0.5 ml/vial）of lyophilized human meningococcal reference serum Men10 were successfully prepared with 2.5%of residual moisture .Reference serum Men10 was sterile and free from contamination by hepatitis B virus , hepatitis C virus , human immunodeficiency virus and syphilis .Provisional IgG concentration of Men 10 to capsular polysaccharide of serogroups A, C, Y and W135 was calibrated by using CDC1992 as the standard.Furthermore, IgG concentrations of both panels of 12 CDC calibration serum samples and 56 LIBP serum samples calibrated by using Men 10 as the standard correlated well with those by using CDC1992 as the standard （r=0.99,P〈

关 键 词：人脑膜炎球菌参考血清 抗体含量 酶联免疫吸附试验 血清杀菌试验 

分 类 号：R392.1[医药卫生—免疫学]