基于逻辑门的邻位核酸荧光传感器同时检测凝血酶和肌红蛋白  

Logic Gate-based Ortho-Nucleic Acid Fluorescence Sensor for Simultaneous Detection of Thrombin and Myoglobin

作　　者：钟子惠 霍冰洋 夏凌 贺锦灿 李攻科 ZHONG Zi-Hui;HUO Bing-Yang;XIA Ling;HE Jin-Can;LI Gong-Ke(School of Chemistry,Sun Yat-sen University,Guangzhou 510006,China;College of Public Health,Guangdong Pharmaceutical University,Guangzhou 510310,China)

机构地区：[1]中山大学化学学院,广州510006 [2]广东药科大学公共卫生学院,广州510310

出　　处：《分析化学》2025年第2期195-203,共9页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金重点项目(No.22134007);广东省自然科学基金项目(No.2024A1515011077)资助。

摘　　要：基于分裂适配体的靶标识别能力和分子逻辑门的智能分析能力,将两种分裂适配体整合到“AND”逻辑门中,构建了一种新型邻位核酸荧光传感器,用于凝血酶(Tob)和肌红蛋白(Myo)的同时检测。当仅存在一种目标物时,信号的响应为单一荧光输出信号,可作为早期低风险判断的评价标准;当两种目标物同时存在时,分裂适配体与目标物结合,形成三元复合物,分别导致首尾的邻位核酸效应,同时触发G-四链体增强硫磺素T荧光信号和Cyanine 3的荧光信号猝灭,可作为早期高风险判断的评价标准。在优化条件下,本方法检测Tob的线性范围为3~200 nmol/L,相关系数为0.9931,检出限(LOD)为0.97 nmol/L;检测Myo的线性范围为6~400 nmol/L,相关系数为0.9933,LOD为2.14 nmol/L。将本方法应用于临床血清样本中Tob和Myo的同时测定,加标回收率分别为85.4%~118.3%和85.8%~119.9%,相对标准偏差均小于6.5%。与标准方法的测定结果相比,本方法的相对误差的范围为-8.8%~5.6%。此外,4例血清样品的逻辑诊断结果为急性心肌梗死高风险2例,低风险2例。本研究提出的“AND”逻辑门邻位核酸荧光传感方法具有选择性高、快速、准确和可同时检测等优点,为急性心肌梗死早期诊断提供了重要参考,也为生物标志物的同时检测提供了一种通用性检测设计思路和平台。Based on the target recognition ability of split aptamer and intelligent analytical capability of molecular logic gate,in this work,two split aptamers were integrated into“AND”logic gate to construct a novel ortho-nucleic acid fluorescence sensor for simultaneous detection of thrombin and myoglobin.When there was one target,the response of the signal was only a single fluorescence output signal,which was used as an evaluation standard for early low-risk judgment.When two targets coexisted,the split aptamer bound to the target to form a ternary complex and led to the head and tail ortho-nucleic acid effect respectively,and triggered the G4 chain to enhance the fluorescence signal of thioflavin T and the fluorescence signal quenching of Cyanine 3,which could be used as an evaluation criterion for early high-risk judgement.Under the optimal conditions,the linear range for detection of thrombin was 3-200 nmol/L,with a correlation coefficient of 0.9931 and a detection limit of 0.97 nmol/L,and the linear range for detection of myoglobin was 6-400 nmol/L,with a correlation coefficient of 0.9933,and a detection limit of 2.14 nmol/L.The method was applied to simultaneous determination of thrombin and myoglobin in clinical serum samples,and the recoveries were 85.4%-118.3%and 85.8%-119.9%,respectively,with relative standard deviations of less than 6.5%.Compared with the standard method,the relative error range was from-8.8%to 5.6%.In addition,the logical diagnosis results of 4 serum samples were high-risk of acute myocardial infarction in 2 cases and low-risk in 2 cases.The″AND″logic gate ortho-nucleic acid fluorescence sensing method showed many advantages such as high selectivity,rapidity,accuracy and simultaneous detection,which offered important reference for early diagnosis of acute myocardial infarction,and also provided a general detection design strategy and platform for simultaneous detection of biomarkers.

关 键 词：逻辑门 分裂适配体 同时检测 凝血酶 肌红蛋白 

分 类 号：R54[医药卫生—心血管疾病]