摘要
玉米赤霉烯酮(Zearalenone,ZEN)是一类真菌毒素,普遍存在于霉变的食物中,对人体的健康带来极大危害。CRISPR-Cas12a是一种RNA引导的核酸内切酶,在基因编辑及生物传感领域受到广泛关注。本文设计了一段ZEN适配体序列ssDNA,该序列能够激活CRISPR-Cas12a的核酸内切酶活性,进而切割固定在电极表面的hp-DNA,从而增大了电极界面电流信号。当ZEN存在时,ssDNA作为ZEN适配体可靶向结合ZEN,从而钝化了CRISPR-Cas12a的酶活性,使得hp-DNA可完好地保留在电极表面,导致电极界面的电流信号降低。基于上述原理,本文构建了一种可用于灵敏检测ZEN的电化学传感方法,该方法对ZEN的检测限达到0.012 ng/mL,可用于实际样品中ZEN的检测。
Zearalenone(ZEN)is a mycotoxin commonly found in moldy foods and poses significant health risks to humans.CRISPR-Cas12a is an RNA-guided endonuclease that has garnered significant attention in gene editing and biosensing applications.In this work,a ZEN aptamer sequence(ssDNA)was designed to activate the endonuclease activity of CRISPR-Cas12a,enabling the cleavage of hairpin DNA(hp-DNA)immobilized on the electrode surface,thereby enhancing the interfacial current signal at the electrode.However,in the presence of ZEN,the ssDNA acts as an aptamer to bind specifically to ZEN,thereby inhibiting the enzymatic activity of CRISPR-Cas12a.This allows the hp-DNA to remain intact on the electrode surface,resulting in a reduced current signal at the electrode interface.Based on this mechanism,an electrochemical sensing method was developed for the sensitive detection of ZEN.The method achieves a detection limit of 0.012 ng/mL and can be applied to the detection of ZEN in beverages.
作者
杨彩萍
童秋霞
魏源
陈薪旭
YANG Caiping;TONG Qiuxia;WEI Yuan;CHEN Xinxu(Fujian Longyan Product Quality Inspection Institute,Longyan 364000;Fujian Longyan Institute of Agricultural Sciences,Longyan 364000;College of Chemistry,Chemical Engineering and Materials Science,Shandong Normal University,Jinan 250014)
出处
《分析科学学报》
北大核心
2025年第4期398-404,共7页
Journal of Analytical Science
基金
福建省龙岩市社会发展科技重点项目(2023LYF9008)
福建省市场监督管理局科技项目(FJMS2023069)。
