CAJ | 학술논문

目的为保证疫苗质量的一致性和可控性,对A、C群脑膜炎奈瑟球菌结合疫苗生产用菌株CMCC(B)29201和CMCC(B)29205株进行毒性和抗原传代稳定性研究,并对30代次菌进行脑腔毒性、免疫原性和产糖质量分析.方法将A、C群脑膜炎奈瑟球菌工作种子批菌种分别连续传代至30代次并收获3、5、10、15、20、25及30代次菌液,小鼠腹腔注射观察各代次毒性,试管凝集试验和间接ELISA测定各代次抗原性.其中30代次菌液进行小鼠脑腔攻击观察是否引起脑组织病理改变,小鼠皮下免疫观察免疫原性,同时进行发酵培养提取荚膜多糖的质量分析.结果CMCC(B)29201株和CMCC(B)29205株工作种子批菌种的半数致死量(LD50)均≥109个/ml,30代次内的LD50也均≥109个/ml,30代次菌液脑腔毒性测定显示均无病理改变;抗原性试管凝集效价均达1∶320,30代次内的试管凝集效价均达1∶320,ELISA几何平均滴度(GMT)分别为1∶4835和1∶3915,且30代次内的ELISA效价分别为1∶4315和1∶3752以上;30代次菌液的血清杀菌抗体均≥1∶32,用第30代次工作种子批菌种生产的A群和C群脑膜炎奈瑟球菌荚膜多糖各项检定指标均达到国家标准.结论 A、C群脑膜炎奈瑟球菌结合疫苗生产用菌株CMCC(B)29201和CMCC(B)29205株毒性低、抗原性和免疫原性良好,连续传代至30代次仍保持较低的毒性和较好的抗原性及免疫原性,纯化的A群和C群脑膜炎奈瑟球菌荚膜多糖质量符合质控要求.
목적 위보증역묘질량적일치성화가공성,대A、C군뇌막염내슬구균결합역묘생산용균주CMCC(B)29201화CMCC(B)29205주진행독성화항원전대은정성연구,병대30대차균진행뇌강독성、면역원성화산당질량분석.방법 장A、C군뇌막염내슬구균공작충자비균충분별련속전대지30대차병수획3、5、10、15、20、25급30대차균액,소서복강주사관찰각대차독성,시관응집시험화간접ELISA측정각대차항원성.기중30대차균액진행소서뇌강공격관찰시부인기뇌조직병리개변,소서피하면역관찰면역원성,동시진행발효배양제취협막다당적질량분석.결과CMCC(B)29201주화CMCC(B)29205주공작충자비균충적반수치사량(LD50)균≥109개/ml,30대차내적LD50야균≥109개/ml,30대차균액뇌강독성측정현시균무병리개변;항원성시관응집효개균체1∶320,30대차내적시관응집효개균체1∶320,ELISA궤하평균적도(GMT)분별위1∶4835화1∶3915,차30대차내적ELISA효개분별위1∶4315화1∶3752이상;30대차균액적혈청살균항체균≥1∶32,용제30대차공작충자비균충생산적A군화C군뇌막염내슬구균협막다당각항검정지표균체도국가표준.결론 A、C군뇌막염내슬구균결합역묘생산용균주CMCC(B)29201화CMCC(B)29205주독성저、항원성화면역원성량호,련속전대지30대차잉보지교저적독성화교호적항원성급면역원성,순화적A군화C군뇌막염내슬구균협막다당질량부합질공요구.
Objective To study the stability of the toxicity,antigenicity and immunogenicity of Neisseria meningitidis serogroup A/C[CMCC(B)29201/29205],and to analyze the quality of the capsular polysaecharide extracted from Neisseria meningitidis.Methods The toxicity of the 3rd,5th,10th,15th,20th,25th and 30th passage of the Neisseria meningitidis was assayed in mice,and the antigenicity of each passage was measured by the tube agglutination test(TAT)and ELISA.The effect of individual 30 passages of Neisseria nveningitidis on the brain tissue and the immunogenicity of each passage were tested in mice,and the capsular polysaccharide was purified and analyzed.Resuits The LD50 of the strains CMCC(B)29201/29205 of each passage was low(≥109/ml),and all the 30 passages of the two strains had no effect on the brain tissue of the mice.The titer of each passage was 1∶320 in TAT and more than 1∶3752 in ELISA.After immunization with individual 30 passages of the Neisseria meningitidis the titers in serum bactericidal assay (SBA)were all more than 1∶32.The capsular polysaccharide purified from Neisseria meningitidis met the quality standard of the requirement.Conclusion The strains of Neisseria meningitidis serogroup A/C used in the manufacture of the meningococcal conjugate vaccine,are stable in the toxicity,antigenicity and immunogenicity.And the capsular polysaccharide has met the quality standard.