| 鲶爱德华氏菌间接酶联免疫快速检测法的建立 |
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| 引用本文： | 李重实,李强,刘海燕,叶仕根,李华.鲶爱德华氏菌间接酶联免疫快速检测法的建立[J].广东海洋大学学报,2010,30(4). |
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| 作者姓名： | 李重实 李强 刘海燕 叶仕根 李华 |
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| 作者单位： | 大连海洋大学,农业部海洋水产增养殖学重点实验室,辽宁,大连,116023 |
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| 基金项目： | 国家十一五科技支撑项目，辽宁省海洋与渔业厅项目，辽宁省教育厅高等学校科研计划，辽宁省自然科学基金 |
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| 摘 要： | 用鲶爱德华氏菌兔抗血清作为一抗,碱性磷酸酶(AP)标记的羊抗兔IgG作为酶标二抗,建立黄颡鱼"红头病"病原菌—鲶爱德华氏菌的间接酶联免疫(ELISA)快速检测法,并优化检测条件。抗原最佳包被浓度为107/mL,一抗工作的最佳稀释度为1∶211,病原菌的检测灵敏度为105/mL,交叉反应实验证明该方法特异性强,与迟钝爱德华氏菌、弧菌等13种标准菌株无交叉。应用上述技术对人工感染发病鱼中分离的优势菌进行检测,结果表明阳性检出率为80%;对自然发病黄颡鱼体内分离获得的20株优势菌检测结果表明,12株菌为鲶爱德华氏菌。
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| 关 键 词： | 鲶爱德华氏菌 酶联免疫吸附实验(ELISA) 黄颡鱼 “红头病” 多克隆抗体 |
Indirect Enzyme-Linked Immunosorbent Assay(ELISA)for Rapid Detection of Edwardsiella ictaluri |
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| LI Zhong-shi,LI Qiang,LIU Hai-yan,YE Shi-gen,LI Hua.Indirect Enzyme-Linked Immunosorbent Assay(ELISA)for Rapid Detection of Edwardsiella ictaluri[J].Journal of Zhanjiang Ocean University,2010,30(4). |
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| Authors: | LI Zhong-shi LI Qiang LIU Hai-yan YE Shi-gen LI Hua |
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| Abstract: | Indirect ELISA for detection of Edwardsiella ictaluri,pathogen of red-head disease of Pelteobagrus fulvidraco was established by using polyclonal antibody against E.ictaluri as primary antibody and alkaline phosphatase conjugated goat-anti-rabbit IgG as second antibody.The optimum coated concentration of the antigen was determined to be 107 cells/mL and the optimum polyclonal antibody concentration was determined to be 1∶211.The lowest concentration of E.ictaluri that can be detected was 105 cells/mL.Cross-reactivity test proved that the method was specificity,and had no cross-reaction with Edwardsiella tarda,Vibrio and other standard strains.With the established method to detect the superior strains isolated from artificial infected fish,the results showed that the positive rate was 80%.12 strains of 20 superior strains isolated from spontaneous diseased P.fulvidraco were identified as E.ictaluri. |
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| Keywords: | Edwardsiella ictaluri Enzyme-Linked Immunosorbent Assay(ELISA) Pelteobagrus fulvidraco "red-head disease" polyclonal antibody |
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