| 草鱼点状产气单胞菌单克隆抗体的制备与双抗原夹心ELISA法的建立 |
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| 引用本文： | 马丽,康洁.草鱼点状产气单胞菌单克隆抗体的制备与双抗原夹心ELISA法的建立[J].贵州农业科学,2012,40(5):138-141. |
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| 作者姓名： | 马丽 康洁 |
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| 作者单位： | 商丘师范学院生命科学系,河南商丘,476000 |
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| 基金项目： | 河南省科技攻关项目，河南省科技厅基础与前沿研究项目 |
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| 摘 要： | 为了建立一种快速、准确能早期检测草鱼肠炎病原菌的方法,以患病草鱼为材料,制备点状产气单胞菌单克隆抗体(McAb),并鉴定其特异性和抗原表位;采用过碘酸钠方法,以辣根过氧化物酶标记单抗,用试管凝集试验测定其活性;建立点状产气单胞菌McAb的双抗原夹心ELISA检测方法,确定包被单抗和酶标单抗的浓度,并测定其灵敏度和特异性。结果:1)获得了2株特异性强和抗原表位不同的单抗,其ELISA效价为1∶16 000。2)辣根过氧化物酶标记的抗体活性为1∶640。3)建立了包被抗体稀释度为1∶800、酶标抗体稀释度为1∶1 600的双抗原夹心ELISA检测方法,该方法与点状产气单胞菌呈强阳性反应,与近似菌无交叉反应,灵敏性为7×103cfu/mL。结论:制备的点状产气单胞菌McAb的效价高、特异性强;建立的双抗原夹心ELISA方法操作简单、灵敏度高、特异性强,可应用于草鱼肠炎病原体的早期快速检测。
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| 关 键 词： | 草鱼 点状产气单胞菌 单克隆抗体 双抗原夹心ELISA |
Preparation of Aeromonas Monoclonal Antibody and Establishment of Double-antigen Sandwich ELISA in Grass Carp |
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| MA Li
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KANG Jie.Preparation of Aeromonas Monoclonal Antibody and Establishment of Double-antigen Sandwich ELISA in Grass Carp[J].Guizhou Agricultural Sciences,2012,40(5):138-141. |
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| Authors: | MA Li KANG Jie |
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| Institution: | (Department of Life Science,Shangqiu Normal University,Shangqiu,Henan 476000,China) |
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| Abstract: | The aeromonas McAb with specificity,epitope and activity was prepared from the silk grass carp and then the McAb double-antigen sandwich ELISA detection method was established for early detecting enteritis pathogens of grass carp.Results:1)two monoclonal antibodies with strong specificity and different epitope were obtained and their ELISA titer was 1∶16 000;2) HRP activity is 1∶640;3)the double-antigen sandwich ELISA detection method with 1∶800 dilution ratio of peridium antibody and 1∶1600 dilution ratio of AbHRP.The method with 7×103 cfu/mL sensitivity shows a strong positive reaction to aeromonas and no cross reaction to the similar bacteria.Conclusion: the prepared aeromonas McAb is of high titer and strong specificity and the established double-antigen sandwich ELISA detection method with simplicity,high sensitivity and strong specificity can be used in early and rapid detection for enteritis pathogen of grass carp. |
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| Keywords: | grass carp aeromonas monoclonal antibody double-antigen sandwich ELISA |
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