正交设计对A群脑膜炎球菌兔抗血清制备条件的筛选  

Screening of preparation of rabbit antiserum against group A Neisseria meningococcus by orthogonal design

作　　者：张荣花 王源 章广玲 ZHANG Ronghua;WANG Yuan;ZHANG Guangling(School of Basic Medical Sciences,North China University of Science and Technology,Tangshan 063000,China)

机构地区：[1]华北理工大学基础医学院

出　　处：《华北理工大学学报（医学版）》2019年第6期421-424,431,共5页Journal of North China University of Science and Technology：Health Sciences Edition

基　　金：华北理工大学科学研究基金资助(编号:Z201607)

摘　　要：①目的应用正交设计筛选A群脑膜炎球菌灭活疫苗的免疫条件,为A群脑膜炎球菌灭活疫苗的应用提供实验依据。②方法将灭活A群脑膜炎球菌浓度调整为1×10^8 CFU/mL、1×10^9 CFU/mL、1×10^10 CFU/mL备用;选择家兔皮内注射、皮下注射、肌肉注射3种方式进行免疫,首次免疫与第二次免疫的时间间隔依次为2、3、4周;根据L9(34)正交设计方案对家兔进行免疫,第二次免疫7天后经家兔颈总动脉采血,分离血清,通过玻片凝集试验和ELISA试验测定兔抗A群脑膜炎球菌血清抗体效价。③结果A群脑膜炎球菌灭活疫苗免疫家兔后,抗原浓度为1×10^9 CFU/mL时血清中抗体的几何效价高于其他两组;皮内注射时抗体的凝集效价高于皮下注射和肌肉注射;免疫间隔时间为3周时,抗体的几何效价最高;差异均具有统计学意义(P<0.05)。A群脑膜炎球菌荚膜多糖与兔抗血清反应的抗体滴度在10^6及以上。④结论免疫剂量为1×10^9 CFU/mL、皮内注射、首次免疫与二次免疫时间间隔为3周时,是家兔的最佳免疫条件,为疫苗血清抗体的制备提供了实验依据。Objective To screening inactivedgroup A Neisseria meningococcu vaccination by orthogonal design,and establish experiment basis for further test.Methods Inactived A group Neisseria meningococcu was diluted into 1x108CFU/mL,1x109 CFU/mL and 1x1010 CFU/mL respectively;Intradermal injection,subcutaneous injection and intramuscularinjection were choosed for antigen entrance and 2,3,4 weeks were the intervals between primary and second immune;Immunization was performed according to the orthogonal design table L9(34).7 days later after the second immune,rabbit carotid artery blood was adopted,and serum was separated,the slide agglutination test and ELISA test were performed to test the specific antibody titer of each groups.Results After inactivedgroup A Neisseria meningococcu vaccine immune rabbit,the geometry meanwas higher than the other two groups at antigen concentration of 1x109 CFU/mL,the geometry mean with Intradermal injection was higher than the other two ways,the geometry mean with 3 weeks interval was higher than the others,and the differences were all significantly.The titers against capsular polysaccharide of group ANeisseria meningococcuwas not less than 106.Conclusion With the dosage of 109 bacteria,intradermal injectionand 3 weeks interval between the first and the second injection were the optimization immune combinationandthe reference for the preparation of serum antibodies induced by vaccines was provided.

关 键 词：A群脑膜炎球菌 正交设计 兔抗血清 

分 类 号：R392.3[医药卫生—免疫学]