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鹿血双抗体夹心间接ELISA检测方法的建立
引用本文:杨舒心,朱明娴,孙长江,谢芳,雷连成.鹿血双抗体夹心间接ELISA检测方法的建立[J].吉林农业大学学报,2010,32(4):440-442.
作者姓名:杨舒心  朱明娴  孙长江  谢芳  雷连成
作者单位:吉林大学畜牧兽医学院,长春,130062
基金项目:吉林省科技发展基金项目 
摘    要:通过制备兔抗鹿血和小鼠抗鹿血多克隆抗体,建立了以兔抗为包被抗体,鼠抗为检测抗体的鹿血双抗体夹心间接ELISA检测方法。采用方阵滴定法确定了抗原、抗体的最佳反应浓度,并对ELISA各反应条件进行优化。结果表明:兔抗最佳包被浓度为1∶8 000,鼠抗的最佳反应浓度为1∶2 000,最佳封闭时间为60 min。该方法对鹿血检测灵敏,可达3×10-4倍稀释度,且对马血、牛血、羊血、猪血的检测结果均为阴性,说明该方法特异性良好。

关 键 词:鹿血  ELISA  多克隆抗体
收稿时间:2009-09-14
修稿时间:2010-01-20

Development of Double-Antibody Sandwich Indirect ELISA to Detect Deer Blood
YANG Shu-xin,ZHU Ming-xian,SUN Chang-jiang,XIE Fang,LEI Lian-cheng.Development of Double-Antibody Sandwich Indirect ELISA to Detect Deer Blood[J].Journal of Jilin Agricultural University,2010,32(4):440-442.
Authors:YANG Shu-xin  ZHU Ming-xian  SUN Chang-jiang  XIE Fang  LEI Lian-cheng
Institution:College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China
Abstract:An double antibody sandwich indirect ELISA was developed fordetectionof deer blood using polyclonal antibody of rabbit as capture antibody and mousespecific polyclonal antibody as detecting antibody. The assay was optimized byphalanx titration to use following antibody dilutions: capture antibody was at 1∶8 000, detecting antibody at 1∶2 000 and the optimal conjugate one hour, respectively. This method had high specificity and sensitivity with 3×10-4and could be used for discerning deer blood distinctly from horse, ox, sheep and pig.
Keywords:ELISA
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