| 柱状黄杆菌间接ELISA快速检测方法的研究 |
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| 引用本文： | 夏君,吴志新,张鹏,熊娟,庞丽娇,陈孝煊.柱状黄杆菌间接ELISA快速检测方法的研究[J].淡水渔业,2009,39(2). |
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| 作者姓名： | 夏君 吴志新 张鹏 熊娟 庞丽娇 陈孝煊 |
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| 作者单位： | 华中农业大学水产学院,武汉,430070 |
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| 基金项目： | 国家科技支撑计划，湖北省科技攻关计划，科技部资助项目 |
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| 摘 要： | 应用间接酶联免疫吸附试验(Indirect-ELISA)技术,建立了快速检测柱状黄杆菌的方法。结果显示:根据棋盘试验,确定最佳抗原包被浓度和免疫血清最佳工作浓度分别为1×106cfu/mL和1∶5000,酶标抗体最佳工作浓度为1∶10000。在该条件下,所建立的间接ELISA能检测出5×104cfu/mL的柱状黄杆菌,且与爱德华氏菌、哈维氏菌弧菌、嗜水气单胞菌、副溶血弧菌等常见鱼类致病菌无交叉反应。结果表明,所建立的间接ELISA可不经细菌培养而直接检测人工感染后草鱼鳃组织中的柱状黄杆菌,该方法对柱状黄杆菌的检测具有快速、敏感、特异、实用的优点。
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| 关 键 词： | 柱状黄杆菌 间接ELISA 检测 |
Indirect ELISA method for detecting Flavobacterium columnaris |
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| XIA Jun,WU Zhi-xin,ZHANG Peng,XIONG Juan,PANG Li-jiao,CHEN Xiao-xuan.Indirect ELISA method for detecting Flavobacterium columnaris[J].Freshwater Fisheries,2009,39(2). |
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| Authors: | XIA Jun WU Zhi-xin ZHANG Peng XIONG Juan PANG Li-jiao CHEN Xiao-xuan |
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| Institution: | XIA Jun,WU Zhi-xin,ZHANG Peng,XIONG Juan,PANG Li-jiao,CHEN Xiao-xuan(Fisheries College of Huazhong Agricultural University,Wuhan 430070) |
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| Abstract: | This study developed an indirect Enzyme-Linked Immunosorbent Assay(Indirect-ELISA) for the rapid detection of Flavobacterium columnaris.The test indicated that the best working dilutions of antigen and antiserum were 1×106 cfu/mL and 1∶5000 separately,which was determined by using checkerboard titration;the best working dilutions of goat anti-rabbit IgG-HRP affinity purified was 1∶10000.In this condition,the detection limit of Indirect-ELISA was approximately 5×104 cfu/mL.Besides,no cross-reaction was observed when Aeromonas hydrophila,Edwarsiella tarda,Vibrioparah aemolyticu,Vibrio harveyi Aeromonas Sobila were applied instead of F.columnaris.Indirect-ELISA can be directly used to detect F.columnaris ingill of grass carp after challenged with F.columnaris.It showed that the Indirec-ELISA is quick,sensitive,specific,effective,and can be used to detect F.columnaris.infection. |
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| Keywords: | Flavobacterium columnaris Indirect-ELISA detection |
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