鱼肠道弧菌单克隆抗体流式细胞术检测技术的研究  

Detection of Bacterial Pathogen Vibrio ichthyoenteri by Monoclonal Antibody and Flow Cytometry

作　　者：邓灯[1] 吴晓春[1] 程顺峰[1] 孙小红[1] 胡春辉[2] 

机构地区：[1]青岛农业大学海洋科学与工程学院,山东青岛266109 [2]青岛农业大学中心实验室,山东青岛266109

出　　处：《水产科学》2015年第2期113-117,共5页Fisheries Science

基　　金：国家自然科学基金资助项目(30901106);国家级大学生创新创业训练计划项目(201310435045);山东省自然科学基金资助项目(ZR2013CQ029)

摘　　要：以抗鱼肠道弧菌单克隆抗体为基础,结合流式细胞术,根据荧光信号强弱比例确定检测鱼肠道弧菌时所需单克隆抗体的最优反应量为200μL,最佳反应时间为45min。在最优反应条件下,抗鱼肠道弧菌单克隆抗体可特异性识别鱼肠道弧菌,阳性荧光信号比例达12.9%;对不同密度的鱼肠道弧菌进行重复性试验,变异系数均在5%以内,说明流式细胞术具有较好的精密度;人工感染健康牙鲆试验表明,流式细胞术检测法能快速从发病鱼的鳃、肝、脾、肾、腹水等部位检测出病原菌,而对照组为阴性;本试验建立的鱼肠道弧菌单克隆抗体流式细胞术检测技术为鱼肠道弧菌的快速检测提供了新方法。In this article,bacterial pathogen Vibrio ichthyoenteri was detected in terms of fluorescence rate by flow cytometry.The best results were found under the reactive conditions of 200μL MAb for 45 min reaction period.The flow cytometry method was shown to provide high reliability of speciality for V.ichthyoenteri in optimal reaction condition,with positive percent of 12.9% and variation coefficient of less than 5%for repeated detection of various concentration samples.Artificial infection test revealed that the V.ichthyoenteri was observed in gill,liver,spleen,kidney and ascites.It is concluded that the established flow cytometry method based on MAb detects V.ichthyoenteri accurately and quickly.

关 键 词：单克隆抗体 流式细胞术 鱼肠道弧菌 

分 类 号：S917[农业科学—水产科学]