| 莱克多巴胺快速检测竞争ELISA试剂盒的研制及其性能测定(英文) |
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| 引用本文： | 张海棠,姜金庆,邓瑞广,王自良.莱克多巴胺快速检测竞争ELISA试剂盒的研制及其性能测定(英文)[J].农业科学与技术,2008,9(5):124-129. |
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| 作者姓名： | 张海棠 姜金庆 邓瑞广 王自良 |
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| 作者单位： | 河南科技学院动物科学学院;河南省动物免疫学重点实验室 |
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| 基金项目： | 国家科技支撑计划重大项目
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| 摘 要： | Mixed anhydride(MA)was used to conjugate ractopamine(RAC)to BSA and obtained artificial antigen BSA-RAC identified by UV and SDS-PAGE.Balb/c mice were immunized with BSA-RAC and hybridoma lines that secrete RAC monoclonal antibody(mAb)were generated with cell fusion.A ciELISA kit for detection of RAC(RAC-Kit)was developed with RAC mAb and its performance were tested.The results indicated that BSA-RAC was successfully synthesized and its conjugation ratio of RAC to BSA was about 24.5∶1.Three hybridoma lines were filtered and the best one was 4D8-3E11,its affinity constant(Ka)was 1.65×1010 L/mol.The limit of detection of RAC-Kit was 0.5 ng/ml and its detection range was 0.5-184 ng/ml.The mean recoveries of RAC spiked in feed were 85.6% and in swine urine were 88.6%.The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were below 15%.It had 9.4% cross-reactivity(CR%)to dobutamine and little or no CR to other compounds.The validity of RAC-Kit in 4 ℃ was in 180 d.
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| 关 键 词： | Ractopamine Artificial antigen Monoclonal antibody Competitive ELISA Rapid test kit |
Development and Performance Measurement of Rapid Detection ciELISA Kit for Ractopamine |
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| ZHANG Hai-tang,JIANG Jin-qing,DENG Riu-guang,WANG Zi-liang.Development and Performance Measurement of Rapid Detection ciELISA Kit for Ractopamine[J].Agricultural Science ＆ Technology,2008,9(5):124-129. |
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| Authors: | ZHANG Hai-tang JIANG Jin-qing DENG Riu-guang WANG Zi-liang |
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| Institution: | 1.College of Animal Science,Henan Institute of Science and Technology,Xinxiang 453003;2.Henan Provincial Key Laboratory for Animal Immunology,Zhengzhou 450002 |
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| Abstract: | Mixed anhydride(MA) was used to conjugate ractopamine(RAC)to BSA and obtained artificial antigen BSA-RAC identified by UV and SDS -PAGE. Balb/c mice were immunized with BSA-RAC and hybridoma lines that secrete RAC monoclonal antibody(mAb) were generated with cell fu-sion. A ciELISA kit for detection of RAC (RAC-Kit) was developed with RAC mAb and its performance were tested. The results indicated that BSA-RAC was successfully synthesized and its conjugation ratio of RAC to BSA was about 24.5:1. Three hybridoma lines were filtered and the best one was 4D8-3E11, its affinity constant (Ka) was 1.65×1010 L/mol. The limit of detection of RAC-Kit was 0.5 ng/ml and its detection range was 0.5-184 ng/ml. The mean recoveries of RAC spiked in feed were 85.6% and in swine urine were 88.6%. The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were below 15%. It had 9.4% cross-reactivity (CR%) to dobutamine and little or no CR to other com-pounds. The validity of RAC-Kit in 4℃ was in 180 d. |
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| Keywords: | Ractopamine Artificial antigen Monoclonal antibody Competitive ELISA Rapid test kit |
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