抗沙门氏菌O8因子单克隆抗体的研制及初步应用  

Preparation and Preliminary Application of Monoclonal Antibodies Against O8 Antigen of Salmonella

作　　者：李珊珊[1] 张志龙 刘丽[1] 王玉金[1] 杜迅[1] 胡宜亮[1] 周伏忠[1] 杨书豪[1] LI Shanshan;ZHANGZhilong;LIU Li;WANGYujin;DU Xun;HU Yiliang;ZHOU Fuzhong;YANG Shuhao(Biological Institute Co.Ltd.,Henan Academy of Sciences,Zhengzhou 450008,China)

机构地区：[1]河南省科学院生物研究所有限公司,郑州450008

出　　处：《河南科学》2018年第6期848-852,共5页Henan Science

基　　金：河南省重点科技攻关项目(152102310126)

摘　　要：制备抗沙门氏菌O8因子单克隆抗体,建立C2、C3亚群沙门氏菌酶联免疫吸附分析检测方法.用加热灭活的纽波特沙门氏菌免疫BALB/c小鼠,通过细胞融合建立分泌抗体的杂交瘤细胞株.用ELISA法检测及鉴定单克隆抗体,酶联免疫吸附分析法初步判断抗体应用于检测试剂的可能性.得到4株与沙门氏菌C2、C3亚群菌株发生反应的单克隆抗体细胞株,分别命名为6F2、7A2、8D8和8D9;4株抗体与肠道正常细菌和常见的致病细菌无交叉反应;单克隆抗体免疫球蛋白类型分别为小鼠IgG_1、IgG_3、IgG_(2b)、IgM,轻链均为kappa;用单克隆抗体7A2和8D8研制的酶联免疫吸附分析试剂盒,对沙门氏菌C2亚群的纽波特沙门氏菌、波那雷恩沙门氏菌和C3亚群的肯塔基沙门氏菌的最低检出值均可达到10~5cfu/m L.获得的单克隆抗体具有较高的特异性,有可能应用于生产检测C2和C3亚群沙门氏菌的酶联免疫吸附分析试剂盒.In order to prepare monoclonal antibodies against O8 antigen of Salmonella and establish enzyme-linked immunosorbent assay for Salmonella C2 and C3 sub-group,the spleen cells from BALB/c mice immunized with heat-killed S. newport were fused with meyloma cells SP2/0. The monoclonal antibodies were detected and identified by ELISA. Possibility of the monoclonal antibodies to detect C2 and C3 sub-group of Salmonella were preliminarily determined by enzyme-linked immunosorbent assay. The result showed that four monoclonal antibody reacted with Salmonella C2 and C3 sub-group were named as 6F2,7A2,8D8 and 8D9 respectively. There was no cross-reaction between the four antibodies and intestinal normal bacteria and common pathogenic bacteria;Immunoglobulin isotype were mouse IgG_1,IgG_3,IgG_（2b） and IgM respectively. The minimum detectable values for S. newport,S. bonariensis and S.kentucky were 10~5 cells/m L by enzyme-linked immunosorbent assay made of monoclonal antibody 7 A2 and 8 D8. The monoclonal antibodies obtained have higher specificity and may be used to produce enzyme-linked immunosorbent assay for detecting C2 and C3 sub-group of Salmonella.

关 键 词：沙门氏菌 O8因子 单克隆抗体 酶联免疫吸附分析 

分 类 号：O939.93[理学] R446.61[医药卫生—诊断学]