Direct detection and quantification of microRNAs.
Direct detection and quantification of microRNAs.
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DOI：
10.1016/j.ab.2009.01.011
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发表时间：
2009-04-01
影响因子：
2.9
通讯作者：
Deo SK
中科院分区：
文献类型：
--
作者：
Hunt EA;Goulding AM;Deo SK
最近发现的 microRNA (miRNA) 是一类相对较新的约 22 个核苷酸 RNA，其强大的调节性质使其成为当今生化和医学研究的主要焦点。 miRNA 表达模式与癌症以及其他疾病发病之间的关系，让我们看到了 miRNA 作为疾病生物标志物或药物靶点的潜力，使其成为主要研究热点。它们在医学上的光明前景取决于提高我们对其复杂调控机制的科学理解。在分析化学领域，与 miRNA 相关的主要挑战是其检测。它们的尺寸极小，细胞浓度低，为获得可靠的结果带来了许多挑战。目前该领域的评论主要集中在微阵列、PCR 和 Northern blotting 程序的改进上，以使其适用于 miRNA 检测。虽然这些方法非常强大并且被接受为当前标准，但它们通常非常费力、半定量，并且通常需要昂贵的成像设备和/或放射性/有毒标签。本综述旨在重点介绍 miRNA 检测和定量方面的新兴技术，这些技术与当前标准相比表现出卓越的灵活性和适应性，以及匹配或更高的灵敏度。具体来说，本综述将涵盖比色、荧光、生物发光、酶和电化学方法，这些方法大大降低了程序复杂性和总体操作费用，从而提高了该研究领域的可及性。介绍并讨论了这些方法相对于当前标准方法的改进以及阻碍其被接受为标准程序的潜在复杂性。这些新方法通过采用基于酶的信号放大、使用 PNA 捕获探针增强杂交条件、高度灵敏和灵活的光谱形式以及极其灵敏的电催化纳米系统等方法，解决了与 miRNA 检测相关的许多问题。
The recent discovery of the potent regulatory nature of microRNAs (miRNAs), a relatively new class of approximately 22 nucleotide RNAs, has made them a primary focus in today’s biochemical and medical research. The relationship between miRNA expression patterns and the onset of cancer, as well as other diseases, has glimpsed the potential of miRNAs as disease biomarkers or drug targets, making them a primary research focus. Their promising future in medicine is hinged upon improving our scientific understanding of their intricate regulatory mechanisms. In the realm of analytical chemistry, the main challenge associated with miRNA is its detection. Their extremely small size and low cellular concentration poses many challenges for achieving reliable results. Current reviews in this area have focused on adaptations to microarray, PCR, and Northern blotting procedures to make them suitable for miRNA detection. While these are extremely powerful methods and accepted as the current standards, they are typically very laborious, semi-quantitative, and often require expensive imaging equipment and/or radioactive/toxic labels. This review aims to highlight emerging techniques in miRNA detection and quantification that exhibit superior flexibility and adaptability as well as matched or increased sensitivity in comparison to the current standards. Specifically, this review will cover colorimetric, fluorescence, bioluminescence, enzyme, and electrochemical based methods, which drastically reduce procedural complexity and overall expense of operation thereby increasing the accessibility of this field of research. The methods are presented and discussed as to their improvements over current standard methods as well as their potential complications preventing acceptance as standard procedures. These new methods have addressed the many of the problems associated with miRNA detection through the employment of enzyme-based signal amplification, enhanced hybridization conditions using PNA capture probes, highly sensitive and flexible forms of spectroscopy, and extremely responsive electrocatalytic nanosystems, among other approaches.