| 柱状黄杆菌双抗体夹心ELISA检测方法的建立 |
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| 引用本文： | 吕娜,殷晓平,张虹茜,等.柱状黄杆菌双抗体夹心ELISA检测方法的建立[J].中国畜牧兽医,2014,41(3):19-23. |
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| 作者姓名： | 吕娜 殷晓平 张虹茜 等 |
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| 摘 要： |
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| 收稿时间： | 2013-09-09 |
Establishment of a Double Antibody Sandwich ELISA for the Detection of Flavobacterium columnaris |
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| LV Na,YIN Xiao-ping,ZHANG Hong-xi,ZHANG Mao-lan,SUN Qiang,ZHAO Guo-kun,ZHANG Jie.Establishment of a Double Antibody Sandwich ELISA for the Detection of Flavobacterium columnaris[J].China Animal Husbandry & Veterinary Medicine,2014,41(3):19-23. |
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| Authors: | LV Na YIN Xiao-ping ZHANG Hong-xi ZHANG Mao-lan SUN Qiang ZHAO Guo-kun ZHANG Jie |
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| Institution: | （1. College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China；2. College of Food Science and Engineering, Jilin Agricultural University, Changchun 130118, China） |
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| Abstract: | The aim of this study was to develop a rapid method for the detection of Flavobacterium columnaris based on a double antibody sandwich ELISA （DAS-ELISA）. Purified monoclonal antibody against Flavobacterium columnaris was used as the capture antibody, while polyclonal antibody was used as the detection antibody. The optimal conditions for the ELISA were as follows: monoclonal antibody with 0.08 μg per well was added to coat overnight at 4 ℃; the plate was blocked by 30 g/L bovin serum albumin for 90 min at 37 ℃; incubation concentration of polyclonal antibody was 0.11 μg per well; the incubation time for detection antigen, polyclonal antibody and enzyme labeled antibody was 1 h at 37 ℃ for each; the value of D492 nm was obtained after 15 min coloration. Judging with P/N≥2.1 and D492 nm≥0.776 as positive criteria. This method had no cross reaction with Edwardsiella tarda, E. coli, Aeromonas hydrophila, Vibrio anguillarum, Vibrio alginolyticus, Vibrio parahaemolyticus and Vibrio harveyi. Its minimum detectable limit was 1×103 CFU. Therefore, this study provided a specific and sensitive detection method for Flavobacterium columnaris for the first time. |
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| Keywords: | Flavobacterium columnaris double antibody sandwich ELISA monoclonal antibody |
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