A群脑膜炎球菌兔抗血清的制备及鉴定  

Preparation and identification of rabbit antiserum against group A Neisseria meningococcus

作　　者：刘鹏[1] 张海峰[1] 郭蓉[1] 薛红刚[1] 

机构地区：[1]武汉生物制品研究所有限责任公司,湖北武汉430207

出　　处：《中国生物制品学杂志》2016年第5期523-527,共5页Chinese Journal of Biologicals

基　　金：国家"重大新药创制"专项<疫苗综合性技术研究开发大平台建设>(2013ZX09402302)

摘　　要：目的制备A群脑膜炎球菌兔抗血清内部参考品,并进行鉴定。方法用新鲜培养的A群脑膜炎球菌混悬液按确定的免疫方案免疫日本大耳白兔,免疫完成后,按确定的采血时间点,经颈动脉采血,分离血清,进行凝集试验,观察兔抗血清凝集反应情况;采用免疫双扩散和ELISA法检测兔抗血清效价,同时利用火箭免疫电泳和ELISA法鉴定兔抗血清的血清学特异性。结果确定免疫方案为基础免疫4周后,加强免疫2周;免疫完成后第6天为最佳采血时间点。6份自制A群脑膜炎球菌兔抗血清的凝集试验效价均达到1︰160,与相应的多糖能够产生沉淀线的最大稀释倍数均不低于1︰8,与A群脑膜炎球菌荚膜多糖反应的抗体滴度均在106及以上,与C、Y和W135群脑膜炎球菌荚膜多糖之间无交叉反应。结论自制的A群脑膜炎球菌兔抗血清可作为A群脑膜炎球菌荚膜多糖疫苗的定性和定量检测的内部参考品,同时也为其他多糖类疫苗血清抗体的制备提供了参考。Objective To prepare and identify the in-house reference for rabbit antiserum against group A Neisseria meningococcus.Methods Rabbits were inoculated with freshly cultured suspension of group A Neisseria meningococcus,of which the serum samples were collected at various time points for agglutination test,then determined for titer by double immunodiffusion and ELISA,and for specificity by rocket immunoelectrophoresis and ELISA.Results The schedule for immunization consisted of a primary immunization（3 times a week for 4 weeks）and a boost immunization（twice a week for 2 weeks）.The optimal time point for collection of serum samples was day 6 after the last immunization.All the agglutination titers of prepared six serum samples reached 1 ∶ 160,while the maximum dilution forming precipitation lines with the corresponding polysaccharides were not less than 1 ∶ 8,and the titers against capsular polysaccharide of group A N.meningococcus was not less than 106.No cross reactions with groups C,Y and W135 N.meningococcus were observed.Conclusion The prepared rabbit antiserum against group A N.meningococcus may be used as the in-house reference for qualitative and quantitative determinations of group A N.meningococcus capsular polysaccharide vaccine,which provides a reference for the preparation of serum antibodies induced by other polysaccharide vaccines.

关 键 词：A群脑膜炎球菌 兔抗血清 

分 类 号：R378.15[医药卫生—病原生物学] R392.33[医药卫生—基础医学]