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投稿时间：2020-10-31
投稿时间：2020-10-31
中文摘要: 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4.12×104cfu/mL～3.4×106cfu/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105cfu/mg，具备较好的灵敏性和特异性。
Abstract: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 4.12×104cfu/mL，and linear range was 4.12×104cfu/mL-3.4×106cfu/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 105cfu/mg in aquatic.
keywords: Shigella double antibody sandwich enzyme-linked immunosorbent assay（ELISA） detection method antibody aquatic
文章编号：202104026 中图分类号： 文献标志码：
基金项目: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）
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