摘要
以阪崎克罗诺杆菌标准菌株ATCC 29004免疫BALB/c小鼠,采用杂交瘤细胞技术,经过两次细胞融合共制备出7株分泌单克隆抗体的杂交瘤细胞株(1E9、2A7、2D10、3E5、5B10、6E3、12A2)。其中2A7、2D10、3E5、12A2的效价达到了1∶256 000,6E3的效价为1∶128 000,5B10的效价为1∶64 000,1E9的效价为1∶32 000。亚型鉴定结果显示1E9的亚型为Ig G1,6E3的亚型为Ig G2b,其余抗体2A7、2D10、3E5、5B10、12A2的亚型均为IgG2a。对7株抗体进行特异性检测结果显示,2A7能与3株阪崎克罗诺杆菌(ATCC 29004、ATCC 29544、ATCC 12868)都结合,1E9能与其中两株(ATCC 29004、ATCC 12868)结合,且与其他11株类似菌和常见致病菌交叉反应结果显示7个抗体均有良好的特异性。用2A7建立的间接竞争酶联免疫吸附测定法对阪崎克罗诺杆菌的检测灵敏度可达到10~6 CFU/m L。
BALB/c mice were immunized with Cronobacter sakazakii type strain ATCC29004.Seven strains(1E9,2A7,2D10,3E5,5B10,6E3,and12A2)of hybridoma cell lines that secreted monoclonal antibodies were prepared by two cycles of cell fusion using hybridoma technique.The titers of2A7,2D10,3E5and12A2reached1:256000,the titer of6E3was1:128000,the titer of5B10was1:64000and the titer of1E9was1:32000.The results of subtype identification showed that the subtype of1E9was IgG1,the subtype of6E3was IgG2b,and the subtypes of the other antibodies2A7,2D10,3E5,5B10and12A2were all IgG2a.2A7could specifically bind to3strains of Cronobacter sakazakii(ATCC29004,ATCC29544,and ATCC12868),and1E9could specifically bind to two of them(ATCC29004and ATCC12868).The results of crossreaction with11other similar pathogens showed that7antibodies had good specificity.The sensitivity of indirect competitive enzyme linked immunosorbent assay(ELISA)using2A7reached106CFU/mL for the detection of Cronobacter sakazakii.
作者
翟绪昭
曾海娟
王广彬
董庆利
谢曼曼
丁承超
刘武康
王淑娟
李杰
刘箐
ZHAI Xuzhao;ZENG Haijuan;WANG Guangbin;DONG Qingli;XIE Manman;DING Chengchao;LIU Wukang;WANG Shujuan;LI Jie;LIU Qing(School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China;Xuzhou Lüjian-Dairy Beverage Co. Ltd., Xuzhou 221006, China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2017年第20期268-272,共5页
Food Science
基金
上海市科委"科技创新行动计划"长三角科技联合攻关领域项目(15395810900)
徐州绿健乳品饮料有限公司"乳制品生产体系致病菌快速检测"项目(3A15308006)
关键词
阪崎克罗诺杆菌
单克隆抗体
间接竞争ELISA
Cronobacter sakazakii
monoclonal antibodies
indirect competitive enzyme linked immunosorbent assay (ELISA)
