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基于CdSe阳离子交换的玉米赤霉烯酮新型荧光免疫检测方法的建立及应用
引用本文:章先,徐浩瑜,赵鹿如,李可,张晓峰,程昌勇,宋厚辉. 基于CdSe阳离子交换的玉米赤霉烯酮新型荧光免疫检测方法的建立及应用[J]. 菌物学报, 2022, 41(5): 819-829. DOI: 10.13346/j.mycosystema.210421
作者姓名:章先  徐浩瑜  赵鹿如  李可  张晓峰  程昌勇  宋厚辉
作者单位:1 浙江大学医学院,浙江 杭州 3100582 浙江农林大学动物科技学院 动物医学院,浙江 临安 3113003 浙江省畜禽绿色生态健康养殖应用技术研究重点实验室,浙江 临安 3113004 浙江省检验检疫科学技术研究院,浙江 杭州 310012
基金项目:浙江省重点研发计划(2021C02058);国家自然科学基金(32002358);国家级大学生创新创业训练计划项目(202010341040)
摘    要:作为镰刀属真菌的次级代谢产物,玉米赤霉烯酮(zearalenone,ZEN)具有强烈的生殖毒性和免疫毒性,严重威胁动物和人类健康。本研究通过采用羧基修饰的CdSe水溶性量子点(quantum dots,QDs)标记ZEN单克隆抗体,并基于CdSe阳离子交换信号增强原理,建立了ZEN新型荧光免疫检测方法(CdSe QDs-FLISA),检测下限(IC10)和半数抑制率(IC50)分别为0.006 ng/mL和0.17 ng/mL,检测区间(IC20–IC80)为0.01–0.45 ng/mL。与ZEN的结构类似物(α-zearalanol、zearalanone、α-zearalenol、β-zearalenol and β-zearalanol)交叉反应性依次为22.3%、13.1%、6.2%、1.6%和3.9%,与农产品中其他真菌毒素如黄曲霉毒素B1(AFB1)、赭曲霉毒素A(OTA)、呕吐毒素(DON)和伏马毒素B1(FB1)几乎不存在交叉反应。该方法...

关 键 词:真菌毒素  单克隆抗体  量子点  信号增强  定量检测
收稿时间:2021-10-26

Sensitive fluorescence immunoassay for the detection of zearalenone base on signal amplification induced by cation exchange in CdSe quantum dots
ZHANG Xian,XU Haoyu,ZHAO Luru,LI Ke,ZHANG Xiaofeng,CHENG Changyong,SONG Houhui. Sensitive fluorescence immunoassay for the detection of zearalenone base on signal amplification induced by cation exchange in CdSe quantum dots[J]. Mycosystema, 2022, 41(5): 819-829. DOI: 10.13346/j.mycosystema.210421
Authors:ZHANG Xian  XU Haoyu  ZHAO Luru  LI Ke  ZHANG Xiaofeng  CHENG Changyong  SONG Houhui
Affiliation:1 School of Medicine, Zhejiang University, Hangzhou 310058, Zhejiang, China2 College of Veterinary Medicine, College of Animal Science and Technology, Zhejiang A&F University, Lin’an 311300, Zhejiang, China3 Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province, Lin’an 311300, Zhejiang, China4 Zhejiang Academy of Science and Technology for Inspection and Quarantine, Hangzhou 310012, Zhejiang, China
Abstract:Zearalenone (ZEN), an estrogenic and carcinogenic mycotoxin, is produced by Fusarium species, including Fusarium graminearum and Fusarium roseum. ZEN can pose threat to animals and humans through direct or indirect intake, as well as be transmitted to the fetus through placenta. Babies and children are more vulnerable to these effects than adults. A rapid and simple detection technique that is easy to operate and yet possesses a high sensitivity is preferred to monitor the ZEN in various foods. Herein, a fluorescence immunoassay via cation exchange reaction in CdSe quantum dots (CdSe QDs-FLISA) for detection of ZEN in cereal samples was outlined, with the detection range of 0.01-0.45 ng/mL and lower detective limit of 0.006 ng/mL. The Cd/Se QDs-FLISA had low cross-reactivity with the ZEN analogues α-zearalanol, zearalanone, α-zearalenol, β-zearalenol and β-zearalanol (22.3%, 13.1%, 6.2%, 1.6% and 3.9%, respectively), and no cross-reactivity (<0.01%) was observed with other mycotoxins, including aflatoxin B1 (AFB1), ochratoxin A (OTA), deoxynivalenol (DON), and fumonisin B1 (FB1). The average recoveries of the spiked corn samples ranged from 85.6% to 110.7% (CV levels ranged from 5.6% to 10.2%). Commercial contaminated sample ZEN quantification data were consistent with the sample LC-MS/MS detection results. The higher sensitivity and shorter testing time make this new approach an efficient technical support for the detection of trace amounts of ZEN in agriproducts. Additionally, this study also provides a reliable and feasible method for detecting other various targets in food safety monitoring fields.
Keywords:mycotoxin  monoclonal antibody  quantum dots  signal enhance  quantitative detection  
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