基于核酸适配体荧光标记及荧光共振能量转移检测赭曲霉毒素A  

Detection of Ochratoxin A Based on Nucleic Acid Aptamer Fluorescence Labeling and Fluorescence Resonance Energy Transfer

作　　者：刘诺亚 赵新月 张晓萌 宋玉竹[1] 张金阳[1] 韩芹芹[1] LIU Nuoya;ZHAO Xinyue;ZHANG Xiaomeng;SONG Yuzhu;ZHANG Jinyang;HAN Qinqin(Faculty of Life Science and Technology,Kunming University of Science and Technology,Kunming 650500)

机构地区：[1]昆明理工大学生命科学与技术学院,云南昆明650500

出　　处：《分析科学学报》2022年第6期711-715,共5页Journal of Analytical Science

基　　金：国家自然科学基金(No.32160601);云南省基础研究项目(No.202001AT070033)。

摘　　要：本研究基于荧光共振能量转移(Fluorescence Resonance Energy Transfer,FRET)原理,以荧光素(Fluorescein,FAM)标记的核酸适配体(Aptamer)及猝灭基团(Dabycl)标记的互补链(cDNA)为主体,构建了赭曲霉毒素A(Ochratoxin A,OTA)的荧光标记检测方法。该方法对OTA的检测限为0.01μg/mL,OTA浓度在0.01~0.25μg/mL范围内与荧光强度线性关系良好(R^(2)=0.9991),回收率在86.40%~97.50%之间,可用于实际样品的检测。与传统检测方法酶联免疫吸附剂分析相比,本方法具有检测快速、灵敏度高、特异性强等优点,为食品中对人体有害物质的检测提供了一种新思路。In this study,a fluorescence resonance energy transfer(FRET)bioassay was developed to detect ochratoxin A(OTA),the fluorescent labeling of OTA was constructed based on the nucleic acid aptamer labeled by fluorescent(FAM)and the complementary chain(cDNA)labeled by Dabycl.The detection limit of this method is 0.01μg/mL,and the linear relationship is good in the concentration range of 0.01-0.25μg/mL(R^(2)=0.9991).It can be used for the detection of actual samples,and the recovery is between 86.40%and 97.50%.Compared with the traditional detection method of ELISA,this method has the advantages of rapid detection,high sensitivity,strong specificity,and provide a new platform for the detection of harmful molecules in food.

关 键 词：核酸适配体 赭曲霉毒素A 荧光共振能量转移 荧光素标记 

分 类 号：O657.3[理学—分析化学]