Highly-Sensitive Amplification-Free Analysis of Multiple miRNAs by Capillary Electrophoresis

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• 作者：David W. Wegman ; Farhad Ghasemi ; Anna Khorshidi ; Burton B. Yang ; Stanley K. Liu ; George M. Yousef ; Sergey N. Krylov
• 刊名：Analytical Chemistry
• 出版年：2015
• 出版时间：January 20, 2015
• 年：2015
• 卷：87
• 期：2
• 页码：1404-1410
• 全文大小：379K
• ISSN：1520-6882

文摘

Sets of deregulated microRNAs (miRNAs), termed miRNA signatures, are promising biomarkers for cancer. Validation of miRNA signatures requires a technique that is accurate, sensitive, capable of detecting multiple miRNAs, fast, robust, and not cost-prohibitive. Direct quantitative analysis of multiple miRNAs (DQAMmiR) is a capillary electrophoresis (CE)-based hybridization assay that was suggested as a methodological platform for validation and clinical use of miRNA signatures. While satisfying the other requirements, DQAMmiR is not sufficiently sensitive to detect low-abundance miRNAs. Here, we solve this problem by combining DQAMmiR with the preconcentration technique, isotachophoresis (ITP). The sensitivity improved 100 times (to 1 pM) allowing us to detect low-abundance miRNAs in an RNA extract. Importantly, ITP-DQAMmiR can be performed in a fully automated mode using a commercial CE instrument making it suitable for practical applications.