Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen , Thorvaldsensvej 40, DK-1871 Frederiksberg C, Denmark.
Anal Chem. 2014 Jul 15;86(14):6823-6. doi: 10.1021/ac5017166. Epub 2014 Jul 3.
In the past decade, microRNAs (miRNAs) have drawn increasing attention due to their role in regulation of gene expression. Especially, their potential as biomarkers in disease diagnostics has motivated miRNA research, including the development of simple, accurate, and sensitive detection methods. The narrow size range of miRNAs (20-24 nucleotides) combined with the chemical properties of conventional reporter tags has hampered the development of multiplexed miRNA assays. In this study, we have used lanthanide-labeled DNA probes for the detection of miRNAs on membranes using laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS). Three miRNAs from Arabidopsis thaliana were analyzed simultaneously with high specificity, and the sensitivity of the method was comparable to radioactive detection (low femtomol range). The perspective of the developed method is highly multiplexed and quantitative miRNA analysis with high specificity and sensitivity.
在过去的十年中,由于 microRNAs (miRNAs) 在基因表达调控中的作用,它们受到了越来越多的关注。特别是,它们作为疾病诊断生物标志物的潜力激发了 miRNA 研究,包括开发简单、准确和敏感的检测方法。miRNAs(20-24 个核苷酸)的狭窄尺寸范围加上常规报告标签的化学性质,阻碍了多重 miRNA 分析的发展。在这项研究中,我们使用镧系元素标记的 DNA 探针,通过激光烧蚀电感耦合等离子体质谱法 (LA-ICPMS) 在膜上检测 miRNA。同时分析了拟南芥中的 3 种 miRNA,具有很高的特异性,该方法的灵敏度与放射性检测相当(低 femtomol 范围)。该方法具有高度的多重性和定量 miRNA 分析的潜力,具有很高的特异性和灵敏度。