HIV-1电化学活性-非活性开关分子信标的构建及验证  

The construction and confirmation of HIV-1 electrochemically active-inactive switching molecular beacon

作　　者：李博[1] 顾大勇[2] 司徒博[1] 李政良[2] 颜晓慧[3] 刘芹兰[1] 郑磊[1] 王前[1] 

机构地区：[1]南方医科大学南方医院检验医学科(系)临床快速检验实验室,广东广州510515 [2]深圳国际旅行卫生保健中心,广东深圳518033 [3]南方医科大学南方医院临床实验研究中心,广东广州510515

出　　处：《检验医学》2012年第5期408-411,共4页Laboratory Medicine

基　　金：广东省科技计划国际合作项目(2008A050200006);广东省科技计划项目(2010A030300006);广东省自然基金项目(S2011010003840);国家质检总局科技计划项目(2011IK142)

摘　　要：目的构建人类免疫缺陷病毒1型(HIV-1)电化学活性-非活性开关分子信标(CAs-MB)并验证其结构及功能。方法合成并纯化针对HIV-1 gag基因保守区序列的CAs-MB,用傅里叶变换红外光谱(FT-IR)和循环伏安法(CV)验证其结构,用差分脉冲伏安法(DPV)信号强度评价CAs-MB的识别功能。结果胭脂红酸(CA)单体与CAs-MB FT-IR图的变化证明CA已标记于MB末端,CV图证明CAs-MB电化学活性ON和OFF状态的存在。DPV结果证实其能够在电化学平台上实现对目标序列的特异性识别,且能够区分一个碱基的差异。结论 CAs-MB作为HIV-1 gag基因的特异性识别元件有望用于构建新型HIV-1电化学基因传感器。Objective To construct human immunodeficiency virus-1 （HIV-1） electrochemically active-inactive switching molecular beacon （CAs-MB） and confirm its structure and function. Methods CAs-MB targeted conserved region of HIV-1 gag gene was prepared and purified. The structure was confirmed by Fourier transform-infrared spectroscopy （FF-IR） and cyclic voltmnmetry （CV）, while its function of recognizing targets was evaluated by differential pulse vohammetry （DPV）. Results The variations of FF-IR spectra of free carminicacid （CA） and CAs-MB indicated that CA had covalently linked onto the ends of MB. The results of CV proved that both ON and OFF statuses of CAs-MB were existing. DPV results illustrated that CAs-MB was effective in signaling the presence of complementary target and discriminating targets in electrochemical gene biosensor that differ by a single nucleotide. Conclusions CAs-MB opens a new detective technique and can be expanded to electrochemical gene biosensor to perform HIV-1 gag gene specificity detection.

关 键 词：人类免疫缺陷病毒1型 分子信标 电化学基因传感器 

分 类 号：R33[医药卫生—人体生理学]