生物技术通报 ›› 2019, Vol. 35 ›› Issue (3): 194-202.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0809

• 技术与方法 • 上一篇    下一篇

功能核酸荧光免标记型定量统一化检测技术的研究进展

肖冰1, 刘榜2, 罗云波1,3, 黄昆仑1,3, 张园1, 李夏莹4, 张秀杰4, 许文涛1,3, 周翔2   

  1. 1. 中国农业大学食品科学与营养工程学院,北京 100083;
    2. 华中农业大学农业动物遗传育种与繁殖教育部重点实验室,武汉 430070;
    3. 农业农村部农业转基因生物安全评价(食用)重点实验室,北京 100083;
    4.农业农村部科技发展中心,北京 100122
  • 收稿日期:2018-09-19 出版日期:2019-03-26 发布日期:2019-04-03
  • 作者简介:肖冰,女,硕士,研究方向:食品生物技术;E-mail:17319374552@163.com
  • 基金资助:
    国家自然科学基金项目(31671922)

Research Progress in Quantitative and Unitive Detecting Technologies of Functional Nucleic Acid and Label-Free Fluorescence

XIAO Bing1, LIU Bang2, LUO Yun-bo1,3, HUANG Kun-lun1,3, ZHANG Yuan1, LI Xia-ying4, ZHANG Xiu-jie4, XU Wen-tao1,3, ZHOU Xiang2   

  1. 1. College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083;
    2. Key Laboratory of Agricultural Animal Genetics,Breeding and Reproduction of the Ministry of education,Huazhong Agricultural University,Wuhan 430070;
    3. Key Laboratory of Safety Assessment of Genetically Modified Organism(Food Safety),the Ministry of Agriculture and Rural Affairs of P.R. China,Beijing 100083;
    4. Department Center for Science and Technology,the Ministry of Agriculture and Rural Affairs of P. R.China,Beijing 100122
  • Received:2018-09-19 Published:2019-03-26 Online:2019-04-03

摘要: 功能核酸是一类具有特定空间构象、执行特异生物功能的天然或者人工核酸序列,具有易于修饰、价格低廉、稳定性高、特异性强等优势,其搭载免标记荧光传感系统后,功能核酸可起到将多种靶物质统一转为较为稳定的核酸信号,以及通过核酸扩增对检测信号进行扩增等重要作用。而免标记荧光传感技术可以在检测中免去荧光标记时荧光、猝灭基团的筛选和标记过程的繁琐和成本,同时可保证与核酸的特异或非特异性结合后产生荧光信号变化。通过两种技术的优势结合,检测的灵敏度、实时性可进一步提高,逐渐被广泛应用于环境污染物检测、食品风险因子检测、疾病诊断等多个领域。首先明确功能核酸与荧光定量检测技术等相关概念,详细阐述了几种重要的荧光物质的特点以及其与功能核酸的分子识别、作用方式与发光机制,接着紧紧围绕该种传感技术的几个特点,从功能核酸荧光免标记型定量统一化检测技术与其实际应用角度进行分类介绍与评价对比,最后就功能核酸荧光免标记型定量统一化检测技术在多种领域的检测分析中的研究意义以及存在的问题进行讨论,并对未来的发展与应用作出展望。

关键词: 功能核酸, 免标记荧光, 定量检测, 生物传感器

Abstract: Functional nucleic acid is a natural or an artificial nucleic acid sequence with specific spatial conformation and biological function. It has advantages such as easy method to modify,low cost,high stability and strong specificity. After combined with label-free fluorescence biosensors,the functional nucleic acid performs the function of transferring all different kinds of targets to stable signals of nucleic acid and amplifying signals by the amplification of nucleic acid. In addition,the complexity of process and cost from labeling and screening of fluorescence,quenching groups in label-fluorescence can be avoided by the technology of label-free fluorescence,while the fluorescence signal transfer can be ensured by the specific or unspecific combination of nucleic acid and fluorescence. Combining the advantages of 2 technologies,the sensitivity and real-time capability can be enhanced,and the functional nucleic acid-based label-free fluorescence biosensors have been widely applied in the detection of environmental pollutants,risk factors of food safety,disease diagnosis,etc. Firstly,we define the concepts of functional nucleic acid and quantitative fluorescence detecting technology,and elaborate in detail the features of key fluorescent materials,as well as their biological molecular recognitions,reaction principles,and luminous models with functional nucleic acid. Then,we describe,assesse,and compare the quantitative and unitive detecting technologies of functional nucleic acid and label-free fluorescence and their applications according to their features. Finally,we discuss the research significance and issues of the quantitative and unitive detecting technologies of functional nucleic acid and label-free fluorescence in detections,and prospect the future development and application.

Key words: functional nucleic acid, label-free fluorescence, quantitative detection, biosensor