HIV-1电化学活性-非活性开关分子信标的构建及验证

检验医学 ›› 2012, Vol. 27 ›› Issue (5): 408-411.

HIV-1电化学活性-非活性开关分子信标的构建及验证

李博

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

收稿日期:2012-05-10 出版日期:2012-05-30 发布日期:2012-05-10
通讯作者: 郑　磊，联系电话：020-62787681。
作者简介:李　博，女，1987年生，硕士，主要从事电化学基因传感器及其临床应用研究。
基金资助:
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）

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

1. Laboratory of Point-of-care Testing，Clinical Laboratory Center，Nanfang Hospital，Southern Medical University，Guangdong Guangzhou 510515，China；2.Shenzhen International Travel Health Care Center， Guangdong Shenzhen 518033，China;3. Research Center of Clinical Medicine， Nanfang Hospital， Southern Medical University, Guangdong Guangzhou 510515, China

Received:2012-05-10 Online:2012-05-30 Published:2012-05-10

摘要/Abstract

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。

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

Abstract: 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 (FT-IR) and cyclic voltammetry (CV), while its function of recognizing targets was evaluated by differential pulse voltammetry (DPV). Results　The variations of FT-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.

Key words: Human immunodeficiency virus-1, Molecular beacon, Electrochemical gene biosensor

李　博;顾大勇;司徒博;李政良;颜晓慧;刘芹兰;郑　磊;王　前. HIV-1电化学活性-非活性开关分子信标的构建及验证[J]. 检验医学, 2012, 27(5): 408-411.

LI Bo;GU Dayong;SITU Bo;LI Zhengliang;YAN Xiaohui;LIU Qinlan;ZHENG Lei;WANG Qian. The construction and confirmation of HIV-1 electrochemically active–inactive switching molecular beacon[J]. , 2012, 27(5): 408-411.

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