CRISPR-Cas12a信号放大策略与便携式血糖仪耦合定量检测黄曲霉毒素B1  

Quantitative detection of aflatoxin B1 by CRISPR-Cas12a signal amplification strategy coupled with portable glucose meter

作　　者：尹会 刘素君[1] 李杨 贺范祥 周媛媛 李娜[1] 舒琴 乔文艳 史铠 YIN Hui;LIU Sujun;LI Yang;HE Fanxiang;ZHOU Yuanyuan;LI Na;SHU Qin;QIAO Wenyan;SHI Kai(College of New Energy Materials and Chemistry,Leshan Normal University,Leshan 614000,China)

机构地区：[1]乐山师范学院新能源材料与化学学院,乐山614000

出　　处：《分析试验室》2024年第3期301-306,共6页Chinese Journal of Analysis Laboratory

基　　金：国家自然科学基金(22204066);四川省科技计划(2022JDRC0126);大学生创新创业训练计划项目(202210649021)资助;。

摘　　要：建立了CRISPR-Cas12a与便携式血糖仪耦合定量检测黄曲霉毒素B1(AFB1)的方法。体系中AFB1能够激活Cas12a的反式切割活性,激活后的Cas12a切割电极上蔗糖酶修饰的发夹探针,使得蔗糖酶游离到电极表面的溶液中。蔗糖酶催化蔗糖产生可以被血糖仪监测的响应信号,进而实现对AFB1的检测。在浓度0.001~0.1 ng/mL范围内,AFB1浓度与血糖信号呈良好的线性关系,线性方程为S=3.5+229.1c,检出限为0.3 pg/mL。该方法特异性强,适用于实际样品中AFB1的检测。A method for the quantitative detection of aflatoxin B1(AFB1)by coupling CRISPR-Cas12a with a portable glucose meter was established.The presence of AFB1 in the system will activate the trans-cleavage activity of CRISPR-Cas12a to cleave the sucrase modified hairpin DNA on the electrode and release sucrase from the electrode surface into the solution.The released sucrase could catalyze sucrose to produce a response signal that can be monitored by the personal glucose meter to realize the detection of AFB1.The developed biosensor exhibited a good linear relationship in AFB1 concentration range of 0.001-0.1 ng/mL,along with the linear equation of S=3.5+229.1c,with the detection limit of 0.3 pg/mL.This method has highly specificity and can be used for the detection of AFB1 in actual samples.

关 键 词：CRISPR-Cas12a 血糖仪 黄曲霉毒素B1 便携式定量检测 

分 类 号：TS201.6[轻工技术与工程—食品科学]