C型肉毒梭菌肉毒毒素的抗原性分析

【目的】通过对分离纯化的C型肉毒毒素抗原性分析,提供快速鉴定动物因C型肉毒梭菌中毒的理论基础。【方法】对分离纯化的C型肉毒梭菌的肉毒毒素灭活后通过SDS-PAGE测定浓度,确定浓度后与弗氏佐剂乳化后免疫家兔,免疫结束后采家兔血液并分离血清。【结果】将不同稀释倍数的肉毒毒素经SDS-PAGE电泳分离后,转移至HybondNC膜,家兔三免后提取的血清作为一抗,用二抗进行Western blotting检测,大约在98.0和53.0 kDa处出现特异反应带;抗原与抗体在琼脂凝胶上结合时,会形成清晰的沉淀带。【结论】分离纯化的C型肉毒毒素,经免疫后家兔可见:C型肉毒毒素具有免疫原性和反应原性,抗体的效价为1:8。

The Antigenicity Analysis of Botulinum Toxin of Clostridium botulinum Type C

【Objective】The study aims to provide the theoretical basis for quick identification of Clostridium botulinum type C through the separation and purification of type C botulinum toxin antigenic analysis.【Method】The experiments were carried out by separating and purifyinng Clostridium botulinum type C in the blood collected from Freund’s adjuvant emulsified rabbit and the botulinum toxin was inactivated to determine the concentration by SDS-PAGE.The rabbit blood was collected after immunization and the serum was separated.【Result】Different dilutions of botulinum toxin were separated by SDS-PAGE electrophoresis,transferred to HybondNC film,and the three free extracted serum from rabbits was used as primary antibody.The secondary antibody was used for Western blotting analysis.At approximately 98.0 and 53.0 kDa,specific reaction zone appeared;antigens and antibodies in agar gel when combined will form a clear precipitation band.【Conclusion】The immunized rabbits,whose botulinum toxin type C was separated and purified by us,are still there.C botulinum toxin has the characteristics of immunogenicity and reactogenicity,and its antibody titer is 1∶ 8.