应用DNA四面体探针电化学生物传感器检测埃博拉病毒核酸  

An electrochemical biosensor based on a DNA tetrahedral nanostructure probe for the detection of Ebola virus nucleotide

作　　者：王金艳[1,2] 孔文[1] 董世彪[1] 贾雷立[1] 刘超[1] 郝荣章[1] 宋宏彬[1] 

机构地区：[1]军事医学科学院疾病预防控制所 [2]兰州军区68202部队

出　　处：《军事医学》2016年第7期554-557,共4页Military Medical Sciences

基　　金：国家863计划资助项目(2015AA020929);北京市科技新星计划资助项目(Z141107001814071);国家传染病重大专项资助项目(AWS13C007;AWS15J006);军事医学科学院创新基金资助项目(2015CXJJ24)

摘　　要：目的建立一种基于电化学生物传感器的快速检测埃博拉病毒核酸的方法。方法利用一步热变性法自组装成带有固定探针的DNA四面体后,通过Au-S键固定至金电极表面制备成新型核酸探针。与合成的埃博拉病毒核酸序列特异性结合后引入Bio-ss DNA检测序列,通过生物素和链霉亲和素的特异性结合作用引入avidinHRP放大信号,利用计时电流法进行检测。结果该传感器可特异性识别和定量检测人工合成的埃博拉病毒的核酸序列,通过实验条件优化,测定核酸的浓度范围在1.0×10^(-9)~5.0×10^(-6)mol/L呈良好的线性关系,检测下限为5.2×10^(-10)mol/L。结论所构建的DNA四面体探针修饰的电化学生物传感器实现了对埃博拉病毒核酸的灵敏、特异检测。Objective To establish a quick electrochemical biosensor for the detection of nucleic acid of Ebola virus.Methods The DNA tetrahedral nanostructure was self-assembled on gold surface by strong Au-S chemical bonds,leaving the target probe at the top. A biotinylated-ss DNA was introduced as the detection probe by specific binding of the captured target sequence,before avidin-horseradish peroxidase（ HRP） was used as a signal amplifier to transduce amperometric signal through interactions with TMB substrate. Results The results indicated that the nucleotide sequence of Ebola virus could be recognized and detected by the sensor. The linear range for the detection of target DNA was from 1. 0 × 10（-9） to5. 0 × 10（-6）mol / L,and the detection limit was 5. 2 × 10（-10） mol / L. Conclusion The fabricated sensor is demonstrated to be sensitive and specific for the detection of Ebola virus nucleotide.

关 键 词：DNA四面体 电化学生物传感器 埃博拉病毒 

分 类 号：R512.8[医药卫生—内科学]