遗传 ›› 2010, Vol. 32 ›› Issue (1): 31-40.doi: 10.3724/SP.J.1005.2010.00031

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MicroRNA定量检测方法的研究进展

景花1, 宋沁馨2, 周国华1,3   

  1. 1. 中国药科大学生命科学与技术学院, 南京 210009; 2. 中国药科大学药物分析教研室, 南京 210009; 3. 华东医学生物技术研究所, 南京 210002
  • 收稿日期:2009-05-14 修回日期:2009-06-28 出版日期:2010-01-20 发布日期:2010-01-15
  • 通讯作者: 周国华 E-mail:ghzhou@public1.ptt.js.cn
  • 基金资助:

    江苏省自然科学基金项目(编号:BK2008067)资助

Advances in approaches for the quantitative detection of microRNAs

JING Hua1, SONG Qin-Xin2, ZHOU Guo-Hua1, 3   

  1. 1. School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China; 2. Department of Pharmaceutical Analysis, China Pharmaceutical University, Nanjing 210009, China; 3. Huadong Research Institute for Medicine and Biotechnics, Nanjing 210002, China
  • Received:2009-05-14 Revised:2009-06-28 Online:2010-01-20 Published:2010-01-15
  • Contact: ZHOU Guo-Hua E-mail:ghzhou@public1.ptt.js.cn

摘要:

MicroRNA是一类内源性的非编码小分子RNA, 通过下调蛋白编码基因的表达而对不同的细胞发育过程起到重要的调控作用。分析组织或细胞样本中microRNA的表达可为研究这类分子的生物学功能提供重要的信息。近年来, 研究者发展了许多方法检测不同的生理和病理学过程中microRNA的表达差异, 并发现microRNA的异常表达与癌症、神经紊乱和心脏疾病等的发生相关。文章系统地介绍了最新发展的microRNA定量检测方法, 详细阐述了基于探针杂交技术的Northern blotting法、微阵列芯片法、纳米金标记法、桥连同位素标记法, 以及基于扩增技术的定量PCR检测法、滚环扩增法、引物入侵法和新一代大规模高通量测序法等, 并对这些方法的优缺点进行了分析比较。

关键词: MicroRNA, 定量检测;研究进展

Abstract:

MicroRNAs (miRNAs) are a class of endogenous non-coding RNAs that play an important role in the control of developmental process of different cells by negative regulation of protein-coding gene expression. Analyzing miRNA expression in tissues or cells can supply valuable information for investigating the biological function of these molecules. Recently, researchers had proposed a number of approaches for analyzing the differences of miRNA expression among dif-ferent physiological or pathological conditions, and found that aberrant expression of miRNA was related to cancers, neu-rological disorders and heart diseases, etc. This review focuses on newly developed strategies for miRNA quantification, and elucidates in detail the probe-hybridization based methods including Northern blotting, microarray, gold nanoparticle labelling, and splinted ligation with radioactive labels. The amplification-based methods including quantitative PCR, rolling cycle amplification, invader assay, and the next generation sequencing methods were also discussed. The advantages and disadvantages of these methods were compared.

Key words: MicroRNA, quantitative detection;research progress