【摘要】：目的:初步建立以玉米赤霉烯酮(ZEN)抗獨(dú)特型單抗替代玉米赤霉烯酮標(biāo)準(zhǔn)品,檢測ZEN殘留的無毒ELISA定量檢測技術(shù)。方法:以ZEN-BSA為包被原,ZEN單抗(1G4)為反應(yīng)抗體,分別以不同濃度的ZEN毒素或ZEN抗獨(dú)特型單抗1D5(Ab2β-1D5)為競爭物,建立競爭抑制曲線。根據(jù)抑制率在20%~80%間,在相同抑制率條件下,ZEN與1D5間濃度對(duì)應(yīng)關(guān)系,繪制替代標(biāo)準(zhǔn)曲線及對(duì)應(yīng)的濃度轉(zhuǎn)換方程。以ZEN類似毒素代替ZEN作為競爭抗原,檢驗(yàn)方法的特異性;板內(nèi)、板間試驗(yàn)檢驗(yàn)方法的精密度。結(jié)果:初步建立了ZEN無毒ELISA定量檢測技術(shù),1D5與ZEN毒素之間的替代標(biāo)準(zhǔn)曲線方程為:y=1.2846x1.9310(y為ZEN毒素濃度ng/ml,x為抗獨(dú)特型抗體濃度(μg/ml)。檢出限(IC2 0)為1.041 ng/ml,檢測范圍為1.041~25.99 ng/ml。方法的批內(nèi)平均變異系數(shù)為3.88%,批間平均變異系數(shù)為4.96%。方法特異性好,與ZEN類似毒素幾乎無交叉反應(yīng)。結(jié)論:利用ZEN單抗和ZEN抗獨(dú)特型單抗初步建立了ZEN的無毒ELISA定量檢測方法。
[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，

本文編號(hào)：2501988