玉米赤霉烯酮液相芯片定量检测方法的研究  

Quantitative Analysis of Zearalenone Using the Microsphere Array Technology

作　　者：宋慧君[1,2] 王玫[2] 于珂[2] 马惠蕊[2] 王玉坤[2] 曹远银[1] 

机构地区：[1]沈阳农业大学植物免疫研究所,沈阳110161 [2]辽宁出入境检验检疫局,辽宁大连116001

出　　处：《沈阳农业大学学报》2011年第4期479-482,共4页Journal of Shenyang Agricultural University

基　　金：中国质检总局资助项目(2009IK152)

摘　　要：建立玉米赤霉烯酮（ZEN）液相芯片定量检测方法。采用间接竞争法原理和液相芯片技术平台,选用ZEN多克隆抗体对ZEN定量检测方法进行探索。首先将ZEN-BSA与36﹟微球偶联,同时加入待检的游离的ZEN小分子和其标记有生物素的ZEN多克隆抗体,偶联抗原和目标抗原竞争结合生物素标记抗体,通过报告分子SA-PE在532nm波长的激光下,即可检测出报告分子发出的荧光强度,荧光强度与待检ZEN的量成反比。通过优化液相反应体系中ZEN多克隆抗体浓度以及抗原抗体孵育时间,建立了ZEN液相芯片定量检测方法。结果表明：本研究建立的ZEN液相芯片定量检测方法,采用的偶联抗原为ZEN-BSA,偶联量为100μg。ZEN多抗临界饱和浓度为10μg.mL-1,偶联抗原和抗体最佳孵育时间为15h。建立的标准曲线线性方程为y=0.0004x＋0.0013（R2=0.9988）,以国际通用的IC50作为衡量标准,其最低检出浓度为3.25ng.mL-1,线性范围为0.05~10ng.mL-1。建立的ZEN液相芯片检测方法,快速、灵敏、简便、适用于大量样本的检测。To establish the method for quantitative analysis of ZEN using the microsphere array technology platform,the quantitative detection of zearalenone（ZEN） was investigated on the microsphere array technology platform and indirect competition theory.In this research,anti-zearalenone was choosed for the quantitative analysis of ZEN.First,ZEN-BSA was coupled with 36﹟microsphere,then it was added in the 1.5mL centrifuge tube with the free target antigen and the biotin labeled Anti-Zearalenone.Botin labeled anti-zearalenone was competitive binded by the coupling antigen and the free target antigen,and the target antigen was detacted by the report molecule SA-PE which was stimulated fluorescence at 532nm laser light.The fluorescence intensity was inversely proportional to the concentration of the free target antigen.Through optimizing the concentration of anti-zearalenone and the incubation time of antigen and antibody,the quantitative detection of ZEN was established eventually.In this research,the coupling antigen was ZEN-BSA with the coupling quantity 100μg.The critical saturated concentration of anti-zearalenone was 10μg·mL-1,the best incubation time of antigen and antibody was 15h.The linear equation was y=0.0004x＋0.0013（R2=0.9988）.As referring to the international current standard of IC50,the results showed that the minimal detecting concentration was 3.25ng·mL-1,the detection range of the standard curve is 0.05-10ng·mL-1.The quantitative analysis of ZEN using the microsphere array technology was rapid,sensitive,simple,and applicable to mass sample testing.

关 键 词：玉米赤霉烯酮 液相芯片技术 ZEN多抗 定量检测 

分 类 号：S513[农业科学—作物学]