基于催化发夹自组装比率电化学双适体传感器检测乳中四环素  

Ratiometric Electrochemical Mixed Aptamers Sensor Based on Catalytic Hairpin Assembly Detect Tetracycline in Milk

作　　者：侯晓庆 张蕴哲[1] 李子坤[2] 卢鑫[3] 杨倩 张伟[1,4,5] HOU Xiaoqing;ZHANG Yunzhe;LI Zikun;LU Xin;YANG Qian;ZHANG Wei(College of Food Science and Technology,Hebei Agricultural University,Baoding 071000,Hebei;Hebei Agricultural University,Baoding 071000,Hebei;Institute of Technology,Hebei Agricultural University,Cangzhou 061100,Hebei;College of Life Sciences,Hebei Agricultural University,Baoding 071000,Hebei;Hebei Provincial Key Laboratory of Analysis and Control for Zoonoses Microbial,Baoding 071000,Hebei;School of Public Health,Hebei University,Baoding 071000,Hebei)

机构地区：[1]河北农业大学食品科技学院,河北保定071000 [2]河北农业大学,河北保定071000 [3]河北农业大学理工系,河北沧州061100 [4]河北农业大学生命科学学院,河北保定071000 [5]河北省人畜共患病原微生物分析与防控重点实验室,河北保定071000 [6]河北大学公共卫生学院,河北保定071000

出　　处：《中国食品学报》2025年第2期423-432,共10页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金项目(32172288,31371772);河北省自然科学基金重点项目(C2019204342);中央引导地方科技发展资金项目(216Z5501G,226Z5503G,236Z5502G);河北省外专百人计划项目(360-0803-JSN-3YGS);河北省博士后科研项目(B2021005007);河北省重点研发计划项目(18275501D);河北省教育厅科研项目(QN2022073);河北省自然科学基金项目(C2019204284)。

摘　　要：目的:本研究旨在建立一种基于催化发夹自组装(CHA)的比率型电化学混合适体传感器,用于灵敏、准确、稳定地检测四环素(TC)。方法:通过2种混合适配体捕获同一靶标来提高识别能力;通过CHA反应实现靶标的无酶信号放大;采用易于区分峰值的二茂铁(Fc)和亚甲基蓝(MB)作为信号开关分子,标记在DNA探针上,产生比率型的电化学响应信号。结果:该传感器检测TC的线性范围50 fg/mL~100 ng/mL,比2种基于单一适配体的传感器的线性范围宽。检出限(LOD)为77 fg/mL,比2种基于单一适配体的传感器的LOD分别低8.6倍和6.1倍。该传感器对TC具有特异性,重复性评估中的相对标准偏差(RSD)为4.2%,构建的传感器保存9 d后信号响应仍能保持在初始值的92%,并成功用于牛奶中TC的检测。人工加标样品检测结果的加标回收率(87.1%~107.2%)与高效液相色谱(HPLC)(92.1%~100.4%)较为一致。结论:本研究构建的比率型电化学混合适体传感器可用于牛奶中TC的灵敏检测,该方法可为检测各种抗生素提供参考。Purpose:This study aimed to establish a ratiometric electrochemical mixed aptamers sensor based on catalytic hairpin assembly(CHA)for sensitive,accurate,and stable detection of tetracycline(TC).Method:This method improveed recognition ability by capturing the same target through two mixed aptamers.Realize enzyme free signal amplification of the target through CHA reaction,using ferrocene(Fc)and methylene blue(MB),which were easily distinguishable from peak values,as signal switch molecules,labeled on DNA probes to generate proportional electrochemical response signals.Result:The linear range of TC detection by this sensor was from 50 fg/mL to 100 ng/mL,which was wider than the linear range of two single aptamer based sensors.The limit of detection(LOD)was 77 fg/mL,which was 8.6 times and 6.1 times lower than the LOD of two single aptamer based sensors,respectively.This sensor had specificity for TC,with a relative standard deviation(RSD)of 4.2%in repeatability evaluation.After 9 days of storage,the signal response of the constructed sensor could still maintain at 92%of the initial value,and it had been successfully used for the detection of TC in milk.The actual sample detection results had a spiked recovery rate(87.1%-107.2%)that was consistent with high performance liquid chromatography(HPLC)(92.1%-100.4%).Conclusion:The ratiometric electrochemical mixed aptamers sensor constructed in this study could be used for sensitive detection of TC in milk,and this method could provide reference for the detection of various antibiotics.

关 键 词：四环素 混合适配体 催化发夹自组装(CHA) 比率策略 电化学适体传感器 

分 类 号：TP2[自动化与计算机技术—检测技术与自动化装置]