量子点标记荧光免疫法检测花生中黄曲霉毒素B_1  

Fluoroimmunoassay using quantum dots label for detecting of aflatoxin B_1 in peanuts

作　　者：李园园[1,2,3] 李培武[1,2,3,4,5] 张奇[1,2,3,4] 张文[1,3,4,5] 丁小霞[1,2,4,5] 张兆威[1,2,3,4] 

机构地区：[1]中国农业科学院油料作物研究所,湖北武汉430062 [2]农业部油料作物生物学与遗传育种重点实验室,湖北武汉430062 [3]农业部生物毒素检测重点实验室,湖北武汉430062 [4]农业部油料产品质量安全风险评估实验室,湖北武汉430062 [5]农业部油料及制品质量监督检验测试中心,湖北武汉430062

出　　处：《中国油料作物学报》2012年第4期438-442,共5页Chinese Journal of Oil Crop Sciences

基　　金：农业部948重点项目(2010-G1;2011-G5);国家自然科学基金(31171702;31101299);国家农业行业科技专项(201203094;201203037);国家科技支撑计划(2012BAB19B09;2011BAD02D02;2010BAD01B07);湖北省自然科学基金(2011CDB358)

摘　　要：建立了基于量子点标记二抗的间接竞争荧光免疫吸附测定方法(indirect competitive fluorescence-linked immunosorbent assay,cFLISA),并研究检测花生中黄曲霉毒素B1的可行性。采用谷胱甘肽为稳定剂,在水相中直接合成碲化镉(CdTe)量子点,并利用1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)与兔抗鼠二抗进行共价偶联,以黄曲霉毒素B1的单克隆抗体建立cFLISA方法。结果表明,该方法的灵敏度和最低检测限值分别为0.023ng/mL和0.001ng/mL,与传统的有机染料FITC-二抗法比较,灵敏度提高了30倍,花生样品加标0.1、0.05和0.025ng/g,回收率范围在88%～116%之间,变异系数均小于10%。建立的cFLISA方法可以较好的检测花生中黄曲霉毒素B1,并为其它真菌毒素的检测提供参考。System of indirect cFLISA（competitive fluorescence-linked immunosorbent assay） was established and evaluated for detecting of peanut aflatoxin B1（AFB1） using CdTe quantum dots as the fluorescence label coupled with secondary antibody.The glutathione modified water-soluble CdTe QDs was synthesized and covalent coupled with the secondary antibody by 1-ethlyl-3-（3-dimethylaminoproyl） carbodiimide hydrochloride（EDC）.And the monoclonal antibody was used to against AFB1.The 50% inhibition value and the limit of detection（LOD） of the cFLISA were 0.023ng/mL and 0.001ng/mL respectively.Compared to organic dye FITC-secondary antibody,the sensitivity of this method was 30-fold higher than that of the traditional method.Recoveries of cFLISA from different spiked peanut samples were from 88% to 116% with a coefficient of variation less than 10%.Result indicated that cFLISA could be used for the detection of AFB1 in peanuts,and could also be a valuable reference to other mycotoxins application.

关 键 词：碲化镉量子点 荧光免疫测定 黄曲霉毒素B1 花生 

分 类 号：S379.7[农业科学—农产品加工]