布鲁氏杆菌LPS抗原制备及其免疫原性测定

摘要

采用热酚水法提取布鲁氏杆菌脂多糖，分离、纯化并SDS -PAGE银染鉴定。纯化的细菌内毒素免疫小鼠，通过斑点ELISA测定血清抗体效价，检测细菌内毒素免疫原性。采用热酚法能获得较高纯度的细菌内毒素，其免疫原性较高，斑点ELISA测血清效价达到1∶1000000，与全菌免疫原性接近。本研究为制备布鲁氏杆菌单克隆抗体和建立布鲁氏杆菌病快速诊断方法奠定研究基础。%The LPS was prepared from M5 Brucella in hot phenol water .With concentration ,enzymatic hydrolysis and alcohol precipitated to re-move impure protein .Then the purified LPS was identified by SDS -PAGE .The BALB/c mice were immunized with purified LPS as antigen to measure its immunogenicity .The serum antibody titers were detected with DOT -ELISA .The results showed that the LPS was highly purified and its immunogenicity was similar to the Brucella .The results lay the foundation for the preparation of monoclonal antibody and establishment of a rapid diagnostic method for Brucellosis .