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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 the internal reference materials of group A meningococcal rabbit antiserum and to identify them. Methods The Japanese white rabbits were immunized with freshly selected group A meningococcal suspension according to the established immunization protocol. After the immunization was completed, the blood was collected from the carotid artery according to the determined blood sampling time, and the serum was collected for agglutination test. Rabbits Anti-serum agglutination reaction; double immunodiffusion and ELISA detection of rabbit antiserum titers, using rocket immunoelectrophoresis and ELISA to identify the serum-specific anti-rabbit serum. Results After four weeks of immunization based on the immunization protocol, the immunizations were boosted for two weeks and on the sixth day after immunization, the best time for blood sampling was obtained. The titer of agglutination test of 6 homemade A meningococcal rabbit antisera reached 1︰160, and the maximum dilution ratio of the corresponding polysaccharide to the precipitating line was not less than 1︰8, respectively. Compared with group A. meningococcal capsular The polysaccharide reaction antibody titers were 106 and above, and C, Y and W135 group meningococcal capsular polysaccharide no cross-reaction. Conclusion The homemade A meningococcal rabbit antiserum can be used as an internal reference for the qualitative and quantitative detection of group A meningococcal capsular polysaccharide vaccine, and also provide a reference for the preparation of other polysaccharide vaccine serum antibodies.