高等学校化学学报 ›› 2013, Vol. 34 ›› Issue (4): 806-812.doi: 10.7503/cjcu20121120

• 分析化学 • 上一篇    下一篇

新型超支状液晶核酸传感器用于p53突变基因的检测

范玉霞, 谭慧, 留玲微, 吴朝阳, 沈国励, 俞汝勤   

  1. 湖南大学化学生物传感与计量学国家重点实验室, 长沙 410082
  • 收稿日期:2012-12-12 出版日期:2013-04-10 发布日期:2013-03-21
  • 通讯作者: 吴朝阳,男,博士,教授,博士生导师,主要从事电化学与生物传感研究.E-mail:zywu@hnu.edu.cn E-mail:zywu@hnu.edu.cn
  • 基金资助:

    国家自然科学基金(批准号:21175037,21277042,J1210040)和新世纪优秀人才支持计划项目(批准号:NECF-11-0132)资助.

Novel Nucleic Acid Signal Amplification Liquid Crystal Biosensor for Detection of p53 Gene Mutation

FAN Yu-Xia, TAN Hui, LIU Ling-Wei, WU Zhao-Yang, SHEN Guo-Li, YU Ru-Qin   

  1. State Key Laboratory for Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082, China
  • Received:2012-12-12 Online:2013-04-10 Published:2013-03-21

摘要:

基于核酸杂交链式反应影响液晶取向的原理, 构建了一种新型的超支状液晶核酸传感器用于检测p53突变基因. 本文突破传统构建超支状分子的方式, 采用杂交链式反应方法, 以目标序列p53突变基因作为引发剂, 3种不同的发卡探针Hairpin A, Hairpin B和Hairpin C为单体, 在温和的条件下, 通过改变单体的浓度和反应时间自发杂交组装形成尺寸和分子量可控的超支状DNA(branched-like DNA, bDNA). 借助捕获探针将该超支状DNA连接到液晶传感基底表面, 观察液晶分子取向改变前后的光学信号, 实现了p53基因含249密码子突变序列的快速检测. 本方法有望为核酸诊断的发展提供一种新的方法和思路.

关键词: 杂交链式反应, 取向改变, 超支状DNA, 液晶生物传感器

Abstract:

A novel nucleic acid liquid crystal biosensor was constructed for the detection of the p53 mutant gene based on the branched nucleic acid hybridization reaction and the orientational transition of liquid crystal. Herein, a specific target DNA sequence was used as an initiator, the three different hairpin probes as dendritic monomers, and the branched-like DNA with controllable molecular size and weight could be formed by changing the concentrations of the monomers and the reaction time under mild conditions. And then the branched-like DNA was hybridized with the capture DNA sequences on the glass substrate. Making use of unique birefringent properties of liquid crystal molecules, the sensor could get an enhanced polarization response signal, and achieved the rapid and specific detection low to 1 nmol/L mutant DNA strands. This simple and sensitive detection method in the detection of DNA has greater practicality, and is expected to be promoted.

Key words: Hybridization chain reaction, Orientation transition, Branched-like DNA, Liquid crystal biosensor

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