布鲁氏菌LPS单克隆抗体修饰的纳米颗粒制备及电镜观察  

Preparation and electron microscopic observation of nanoparticles modified by Brucella LPS monoclonal antibody

作　　者：李楠[1] 郝镯 祝令伟 纪雪 郭学军 孙洋 田宇飞 郎需龙 LI Nan;HAO Zhuo;ZHU Lingwei;JI Xue;GUO Xuejun;SUN Yang;TIAN Yufei;LANG Xulong(Changchun Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Changchun 130122,China)

机构地区：[1]中国农业科学院长春兽医研究所,长春130122

出　　处：《黑龙江畜牧兽医》2022年第1期78-81,91,共5页Heilongjiang Animal Science And veterinary Medicine

基　　金：“十三五”国家重点研发计划项目(2018YFD0500900;2018YFD0500903)。

摘　　要：为了研究新型纳米材料在免疫电镜技术及病原超微结构观察、定位等方面的应用,试验用羧基量子点微球、时间分辨荧光微球、胶体金颗粒3种纳米材料对布鲁氏菌LPS单克隆抗体进行标记,再与布鲁氏菌病试管凝集试验抗原结合成抗原-抗体免疫复合物,并用电镜观察标记结合效果。结果表明:在电镜下,可观察到布鲁氏菌病试管凝集试验抗原以聚集或单个菌体形式存在。羧基量子点微球、时间分辨荧光微球均以几个或多个颗粒聚集形式存在,其大小、形貌在标记前后并无显著差异;胶体金颗粒则较为均匀地分散在视野中,标记后的胶体金颗粒周围可见抗体晕;标记抗体的微球可与布鲁氏菌病试管凝集试验抗原特异性结合,视野背景干净,菌体呈球杆状,菌体周围吸附了多个纳米颗粒。说明新型纳米材料可用于病原体的免疫电镜技术及微观形态观察、定位等研究。In order to study the application of new nanoparticles in immunoelectron microscopy and pathogen ultrastructure observation and positioning, the experiment used three nanomaterials including carboxyl quantum dot microspheres, time-resolved fluorescent microspheres, colloidal gold particles to label with Brucella LPS monoclonal antibody, which then was combined with the Brucella antigen of tube agglutination test to form an antigen-antibody immune complex, and the combined effect of the label was observed with electron microscope. The results showed that under the electron microscope, it could be observed that the Brucella antigen of tube agglutination test existed in the form of aggregates or single bacteria. Both carboxyl quantum dot microspheres and time-resolved fluorescent microspheres existed in the form of aggregation of several or more particles, and there was no significant difference in the size and morphology of the nanoparticles before and after labeling. The colloidal gold particles were more evenly dispersed in the field of vision, and antibody halos could be seen around the labeled colloidal gold particles. The antibody-labeled microspheres could specifically bind to the Brucella antigen of the agglutination test;the background of the field of view was clean;the bacteria were in the shape of a rod, and a number of nanoparticles were adsorbed around the bacteria. The results suggested that the new nanoparticles could be used in the research of immunoelectron microscopy and microscopic morphological observation and positioning of the pathogens.

关 键 词：布鲁氏菌 透射电子显微镜 免疫标记 量子点微球 时间分辨荧光微球 胶体金 

分 类 号：S852.43[农业科学—基础兽医学]