基于独特型-抗独特型初步建立玉米赤霉烯酮无毒ELISA定量检测技术
发布时间：2019-06-19 02:15
【摘要】：目的:初步建立以玉米赤霉烯酮(ZEN)抗独特型单抗替代玉米赤霉烯酮标准品,检测ZEN残留的无毒ELISA定量检测技术。方法:以ZEN-BSA为包被原,ZEN单抗(1G4)为反应抗体,分别以不同浓度的ZEN毒素或ZEN抗独特型单抗1D5(Ab2β-1D5)为竞争物,建立竞争抑制曲线。根据抑制率在20%~80%间,在相同抑制率条件下,ZEN与1D5间浓度对应关系,绘制替代标准曲线及对应的浓度转换方程。以ZEN类似毒素代替ZEN作为竞争抗原,检验方法的特异性;板内、板间试验检验方法的精密度。结果:初步建立了ZEN无毒ELISA定量检测技术,1D5与ZEN毒素之间的替代标准曲线方程为:y=1.2846x1.9310(y为ZEN毒素浓度ng/ml,x为抗独特型抗体浓度(μg/ml)。检出限(IC2 0)为1.041 ng/ml,检测范围为1.041~25.99 ng/ml。方法的批内平均变异系数为3.88%,批间平均变异系数为4.96%。方法特异性好,与ZEN类似毒素几乎无交叉反应。结论:利用ZEN单抗和ZEN抗独特型单抗初步建立了ZEN的无毒ELISA定量检测方法。
[Abstract]:Objective: to establish a non-toxic ELISA quantitative detection technique for the detection of ZEN residues with zearalenone (ZEN) anti-idiotypic monoclonal antibody instead of zearalenone standard. Methods: ZEN-BSA was used as coating antigen, ZEN monoclonal antibody (1G4) was used as reactive antibody, and different concentrations of ZEN toxin or ZEN anti-idiotypic monoclonal antibody 1D5 (Ab2 尾-1D5) were used as competitors to establish competitive inhibition curve. According to the concentration corresponding relationship between ZEN and 1D5 under the same inhibition rate, the substitution standard curve and the corresponding concentration conversion equation were drawn according to the inhibition rate of 20% 鈮，
本文编号：2501988
[Abstract]:Objective: to establish a non-toxic ELISA quantitative detection technique for the detection of ZEN residues with zearalenone (ZEN) anti-idiotypic monoclonal antibody instead of zearalenone standard. Methods: ZEN-BSA was used as coating antigen, ZEN monoclonal antibody (1G4) was used as reactive antibody, and different concentrations of ZEN toxin or ZEN anti-idiotypic monoclonal antibody 1D5 (Ab2 尾-1D5) were used as competitors to establish competitive inhibition curve. According to the concentration corresponding relationship between ZEN and 1D5 under the same inhibition rate, the substitution standard curve and the corresponding concentration conversion equation were drawn according to the inhibition rate of 20% 鈮，
本文编号：2501988
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