水产品中志贺氏菌双抗夹心ELISA检测方法建立与应用  

Establishment and Application of Double Antibody Sandwich ELISA to Detect Shigella in Aquatic

作　　者：朱芳茜 何扩[1] 张秀媛[1] 李育峰[1] 陈一[1] 王云峰[1] 王硕[2] 赵瑞平[1] ZHU Fang-qian;HE Kuo;ZHANG Xiu-yuan;LI Yu-feng;CHEN Yi;WANG Yun-feng;WANG Shuo;ZHAO Rui-ping(Hebei Key Laboratory of Quality&Safety Analysis-Testing for Agro-Products and Food,Hebei North University,Zhangjiakou 075000,Hebei,China;Nankai University,Tianjin 300000,China)

机构地区：[1]河北省农产品食品质量安全与检测重点实验室,河北北方学院,河北张家口075000 [2]南开大学,天津300000

出　　处：《食品研究与开发》2021年第4期156-160,共5页Food Research and Development

基　　金：河北省引进国外智力项目;河北省农业科技成果转化资金项目(19822801D、20822906D);石家庄市科学技术研究与发展计划项目(191170034A、191170082A);石家庄市引进国外智力项目;河北省自然科学基金项目(C2020405003);石家庄市科技孵化计划项目(191540119A)。

摘　　要：志贺氏菌(Shigella)通称痢疾杆菌,是一类具有传染性强和危害严重的革兰阴性食源性致病菌,广泛污染各类动物源性食品。该研究将志贺氏菌经0.5%福尔马林溶液灭活后,多次免疫雌性大耳兔获得抗志贺氏菌多克隆抗体,以抗志贺氏菌多克隆抗体作为捕获抗体,以辣根过氧化物酶(horseradish peroxidase,HRP)标记抗志贺氏菌多克隆抗体作为检测抗体,建立快速检测志贺氏菌双抗夹心酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)方法。结果表明:建立的双抗夹心ELISA方法检测曲线的线性范围为4.12×104 cfu/mL~3.4×106 cfu/mL,检测限为4.12×104 cfu/mL;用此体系检测大肠埃希氏菌、鼠伤寒沙门氏菌、单增李斯特菌、金黄色葡萄球菌及副溶血弧菌等无交叉反应;水产品实际样品志贺氏菌检出限(limit of detection,LOD)为105 cfu/mg,具备较好的灵敏性和特异性。The Shigella is a kind of gram-negative food-borne pathogenic bacteria with strong infectivity and serious harm,and extensively contaminate all kinds of animal derived food.In this study,polyclonal antibodies were obtained by immunization female big-eared rabbit with Shigella antigen.The sandwich enzyme-linked immunosorbent assay(ELISA)system was established by the horseradish peroxidase conjugated antibody(HRP-antibody)as the detection antibody and the polyclonal antibody as the capture antibody.It showed that the detection limit was 4.12×104 cfu/mL,and linear range was 4.12×104 cfu/mL-3.4×106 cfu/mL.The system was used to detect Escherichia coli,Salmonella enterica,Listeria monocytogene,Staphylococcus aureus and Vibrio parahaemolyticus.The results showed that there were no cross-reactions.The detection limit was 105 cfu/mg in aquatic.

关 键 词：志贺氏菌 双抗夹心 酶联免疫吸附试验(ELISA) 抗体 检测方法 水产品 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程]