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当前位置: 首页 > 文章 > 产志贺毒素大肠杆菌志贺毒素Ⅰ型双抗体夹心ELISA检测方法的建立 畜牧与兽医 2016,48 (8) 18-21
Position: Home > Articles > Establishment of double- antibody sandwich enzyme- linked immunosorbent assay for detection of shiga toxin type Ⅰ in shiga toxin- producing Escherichia coli Animal Husbandry & Veterinary Medicine 2016,48 (8) 18-21

产志贺毒素大肠杆菌志贺毒素Ⅰ型双抗体夹心ELISA检测方法的建立

作  者：
宋路;曾晓燕;史凤娟;黄明明;宋小凯;李祥瑞;焦永军
单  位：
卫生部肠道病原微生物重点实验室江苏省疾病预防控制中心病原微生物研究所;南京农业大学动物医学院
关键词：
产志贺毒素大肠杆菌;Ⅰ型志贺毒素;单克隆抗体;双抗体夹心ELISA
摘  要：
为建立双抗体夹心ELISA法检测产志贺毒素大肠杆菌(STEC)Ⅰ型志贺毒素(Stx1),利用重组Ⅰ型志贺毒素A亚单位(Stx1A)免疫小鼠,制备单克隆抗体,从中筛选出非竞争性的2株单抗,构建双抗体夹心法ELISA检测方法,并对试验样品中的志贺毒素进行检测。结果显示:共获得4株针对Stx1A的单克隆抗体,分别为1H2-G7、2H1-C12、8E7-E6和2F6-F8。当8E7-E6作为包被抗体,而2F6-F8作为检测抗体时,其检测毒素的OD值最高。对样品中的Stx进行检测,表明该方法只有效识别Stx1,对Stx2不识别。结论:成功建立了用于检测Stx1的双抗体夹心ELISA法,为临床上快速诊断STEC感染奠定了基础。
译  名：
Establishment of double- antibody sandwich enzyme- linked immunosorbent assay for detection of shiga toxin type Ⅰ in shiga toxin- producing Escherichia coli
作  者：
SONG Lu;ZENG Xiaoyan;SHI Fengjuan;HUANG Mingming;SONG Xiaokai;LI Xiangrui;JIAO Yongjun;College of Veterinary Medicine,Nanjing Agricultural University;Key Laboratory of Enteric Pathogenic Microbiology,Ministry of Health,Institute of Pathogenic Microbiology,Jiangsu Provincial Center for Disease Prevention and Control;
关键词：
shiga toxin-producing Escherichia coli(STEC);;shiga toxin type Ⅰ(Stx1);;monoclonal antibody(MAb);;double-anti body sandwich ELISA
摘  要：
To establish double- antibody sandwich enzyme- linked immunosorbent assay( ELISA) for detection of shiga toxin type 1( Stx1) of shiga toxin- producing Escherichia coli( STEC),the recombinant shiga toxin type I A subunit( Stx1A) was used to immunize mice to develop monoclonal antibodies( MAbs),and two un- competitive MAbs were screened to set up double- antibody sandwich ELISA.This method was further evaluated by detecting Stx1 from supernatants of STEC cultures. Four strains of hybridomas which can secrete MAbs against Stx1 A,namely 1H2- G7,2H1- C12,8E7- E6 and 2F6- F8,were obtained. The highest OD value could be obtained using 8E7- E6 as coating antibody and 2F6- F8 as detection antibody in double- antibody sandwich ELISA. The ELISA method could detect Stx1 but not Stx2. It suggests that a double- antibody sandwich ELISA for detecting Stx1 was successfully established,which provides a foundation for the clinical diagnosis of STEC infection.

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