金黄色葡萄球菌肠毒素B单克隆抗体制备及免疫酶检测方法的构建  

The development of monoclonal antibodies for staphylococcal enterotoxin B and establishment of immuno-enzymatic method for its detection

作　　者：陈一兵 王雯蕾 张评浒 CHEN Yibing;WANG Wenlei;ZHANG Pinghu(College of Veterinary Medicine,Yangzhou University,Yangzhou,225009,China;Institute of Translational Medicine,Medical College,Yangzhou University,Yangzhou,225009,China)

机构地区：[1]扬州大学动物实验中心,江苏扬州225009 [2]扬州大学医学院转化医学研究院,江苏扬州225009

出　　处：《畜牧与兽医》2025年第5期84-87,共4页Animal Husbandry & Veterinary Medicine

基　　金：扬州大学高端创新人才计划(20180508)。

摘　　要：旨在建立一种免疫酶测定方法,用于定量检测金黄色葡萄球菌肠毒素B。以金黄色葡萄球菌重组肠毒素B免疫BALB/c小鼠,制备鼠源单抗;筛选针对该毒素的配对单抗,构建双抗体夹心ELISA检测体系,在保证检测特异性基础上增加生物素/链霉亲和素放大系统,进一步提升敏感度。通过杂交瘤技术共获得6株针对重组B毒素的单抗克隆,其中由0085(捕获抗体)和0020(检测抗体)构建的双抗体夹心ELISA检测体系表现出良好的特异性,与肠毒素A、C、D、E均无交叉,对肠毒素B的检测敏感度可达0.625 ng/mL。增加生物素/链霉亲和素放大系统后敏感度可提升至0.156 ng/mL。本研究构建的检测方法具有高特异性及高敏感性,具有较强的应用前景。This study was to establish an immuno-enzymatic platform for staphylococcal enterotoxin B detection.Recombinant staphylococcal enterotoxin B(reSEB) was used to immunize mice for monoclonal antibodies development,and two antibody clones were screened to set up an immuno-enzymatic method.Based on its higher specificity,the system was further integrated with biotin/streptavidin to improve its sensitivity.Six antibody clones were obtained and clones 0085 and 0020 were screened to establish a double antibody sandwich ELISA system.This system had a good specificity for SEB,with no cross-binding to SEA,SEC,SED,and SEE.Furthermore,the sensitivity has enhanced from 0.625 to 0.156 ng/mL after biotin/streptavidin modification.The assay system established in this study had higher sensitivity and specificity,and might be used for SEB detection in practice.

关 键 词：金黄色葡萄球菌肠毒素B 单克隆抗体 双抗体夹心ELISA 

分 类 号：S852[农业科学—基础兽医学]