鹿血双抗体夹心间接ELISA检测方法的建立

Development of Double-Antibody Sandwich Indirect ELISA to Detect Deer Blood

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。

An double-antibody sandwich indirect ELISA was developed for detection of deer blood using polyclonal antibody of rabbit as capture antibody and mouse specific polyclonal antibody as detecting antibody. The assay was optimized by phalanx 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^-4 and could be used for discerning deer blood distinctly from horse, ox, sheep and pig.