生物条码凝胶分析法检测 microRNA

Bio-barcode gel assay for microRNA.

机构信息

1] Department of Chemistry, Seoul National University, Seoul 151-747, South Korea [2].

Department of Chemistry, Seoul National University, Seoul 151-747, South Korea.

出版信息

Nat Commun. 2014 Feb 26;5:3367. doi: 10.1038/ncomms4367.

DOI:10.1038/ncomms4367

PMID:24569571

Abstract

MicroRNA has been identified as a potential biomarker because expression level of microRNA is correlated with various cancers. Its detection at low concentrations would be highly beneficial for cancer diagnosis. Here, we develop a new type of a DNA-modified gold nanoparticle-based bio-barcode assay that uses a conventional gel electrophoresis platform and potassium cyanide chemistry and show this assay can detect microRNA at aM levels without enzymatic amplification. It is also shown that single-base-mismatched microRNA can be differentiated from perfectly matched microRNA and the multiplexed detection of various combinations of microRNA sequences is possible with this approach. Finally, differently expressed microRNA levels are selectively detected from cancer cells using the bio-barcode gel assay, and the results are compared with conventional polymerase chain reaction-based results. The method and results shown herein pave the way for practical use of a conventional gel electrophoresis for detecting biomolecules of interest even at aM level without polymerase chain reaction amplification.

摘要

MicroRNA 已被鉴定为一种潜在的生物标志物，因为 microRNA 的表达水平与各种癌症相关。在低浓度下检测它将对癌症诊断非常有益。在这里，我们开发了一种新型的基于 DNA 修饰金纳米粒子的生物条码分析方法，该方法使用常规的凝胶电泳平台和氰化钾化学，并表明该分析方法可以在没有酶扩增的情况下以 aM 水平检测 microRNA。还表明，单碱基错配的 microRNA 可以与完全匹配的 microRNA 区分开来，并且可以通过这种方法对各种 microRNA 序列的组合进行多重检测。最后，使用生物条码凝胶分析从癌细胞中选择性地检测到差异表达的 microRNA 水平，并将结果与传统的基于聚合酶链反应的结果进行比较。本文所示的方法和结果为即使在没有聚合酶链反应扩增的情况下，使用常规凝胶电泳检测感兴趣的生物分子（甚至在 aM 水平）的实际应用铺平了道路。