大肠埃希菌F4菌毛凝集性单抗制备及抗原表位差异

发布时间：2018-03-15 04:20

本文选题：产肠毒素大肠埃希菌F　切入点：凝集性　出处：《扬州大学学报(农业与生命科学版)》2017年01期 　论文类型：期刊论文

【摘要】：采用杂交瘤单克隆抗体制备技术,获得4株能稳定分泌产肠毒素大肠埃希菌(ETEC)F4菌毛特异性抗体的杂交瘤细胞株,分别命名为8EH2、8EH3、9E10、11D4。免疫印迹和凝集反应结果表明:这4株单抗均能与纯化的F4菌毛蛋白发生特异性反应,且与F4大肠埃希菌不同血清型变异株F4ab、F4ac和F4ad发生特异性肉眼可见的凝集反应;而与F18、F6(987P)、F5(K99)产肠毒素大肠埃希菌参考株、分离株以及肠炎沙门氏菌参考株50336、分离株等多种肠杆菌科细菌无任何交叉反应。对单抗亚类的特性检测结果显示,这4株单抗均属于IgG1抗体,且主要识别F4ac的抗原表位,其中,8EH2、8EH3、9E10也能够较好地识别F4ab的抗原表位。结果说明F4菌毛凝集性单克隆抗体为ETEC F4感染的快速特异诊断提供技术基础。
[Abstract]:By using hybridoma monoclonal antibody preparation technique, four hybridoma cell lines were obtained, which could stably secrete the specific antibody against enterotoxigenic Escherichia coli ETEC-F4 fimbriae. The results of immunoblotting and agglutination showed that the four McAbs could react specifically with the purified F4 fimbriae protein. Specific agglutination reaction was observed with different serotype variants of Escherichia coli F4, F4abanF4ac and F4ad, but with F18, F6, 987Pn987, F5K99, the reference strain of Enterotoxin-producing Escherichia coli, F4abanf4ac and F4ad. There was no cross reaction among the isolates, the reference strain of Salmonella enteritidis 50336, and the isolates of Enterobacteriaceae. The characteristics of the McAb subclasses showed that all the McAbs belonged to the IgG1 antibody and mainly recognized the epitopes of F4ac. The antigenic epitopes of F4ab could also be well identified by 8EH _ 2 and 8EH _ 3G _ 9E10. The results indicated that the F4 pili agglutination monoclonal antibody could provide a technical basis for the rapid and specific diagnosis of ETEC F4 infection.
【作者单位】：扬州大学兽医学院/江苏省动物重要疫病与人兽共患病防控协同创新中心;扬州大学动物科学与技术学院;
【基金】：科技部畜禽重大疫病防控与高效安全养殖综合技术研发重点专项(2016YFD0500900) 国家自然科学基金资助项目(31672579、30571374、30771603、31072136、31270171) 江苏高校优势学科建设工程项目(2014-09) 中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室开放基金项目(SKLVBF201706)
【分类号】：S852.4

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