快速检测结核杆菌DNA电化学传感技术的研究  

Research of DNA electrochemical biosensor technology for the rapid detection of tubercle bacillus

作　　者：洪国粦[1,2] 刘银环[1] 潘鸿锦[3] 傅希[1] 章涛[4] 

机构地区：[1]厦门大学附属福州第二医院检验科,福州350007 [2]福建医科大学医学技术与工程学院医学检验系 [3]厦门大学附属福州第二医院胸外科 [4]福建医科大学基础医学院免疫学系

出　　处：《中华实验外科杂志》2012年第11期2286-2288,共3页Chinese Journal of Experimental Surgery

基　　金：福建省自然科学基金资助项目（2012J01428）;福建省医学创新课题（2012-CX-30）;福州市科技计划项目（2011-S-7-3）

摘　　要：目的构建DNA电化学传感器用于快速检测结核杆菌的新方法。方法根据互补序列一段设计捕获探针，另一段设计信号探针，两段探针与互补链结合构建“三明治”电化学传感模式，结合酶的催化作用放大电化学信号，实现对靶序列的高灵敏检测。结果在优化条件下目标链浓度（10nmol／L）检测结果的相对标准偏差（RSD）=5.0％（n=5）；目标DNA在浓度为50-200pmol／L范围内与电流值呈线性相关，回归方程：电流（I）=720．4＋93．6x浓度（c），r=0．9982，检出限为1．6×10^12mol／L。结论此方法构建的传感器具有较高的检测灵敏度与良好的特异性。Objective DNA electrochemical biosensor, as a novel technology, is establised to screen tubercule bacillus. Methods Based on the specific sequence IS6110 of tuberculosis bacilli, cap- ture probe and signal probe are corresponding to the two segments of the complementary target chain, and they are used to form ＂sandwich＂ structure with target sequence. Then horseradish peroxidase （HRP） is added to the ＂sandwich＂ structure to catalyze the reaction of substate and amplific the electrochemical sig- nal for detection of target sequence. Results On optimum conditions, the method was used to parallel de- tect 10 nmol/L target DNA for five times, with relative standard deviation（RSD） 5.0%. The relationship between current and the concentration of the synthetic target complementary strand is linear in the range from 50 pmol/L to 200 pmol/L, with the detection limit of 1.6 ×10^12 mol/L. The equation can be expressed as I（nA） = 720. 4 ＋ 93.6c （ target sequence） , r = 0. 9982. Conclusion The biosensor was estab- lised for the screening of tubercle bacillus with high sensitivity and good selectivity.

关 键 词：结核杆菌 生物传感器 DNA 

分 类 号：R52[医药卫生—内科学]