Multiplexed microRNA detection by capillary electrophoresis with laser-induced fluorescence

Ruei Min Jiang, Yu Sun Chang, Shu Jen Chen, Jian Hung Chen, Hua Chien Chen, Po Ling Chang*

*此作品的通信作者

研究成果: 期刊稿件文章同行評審

30 引文 斯高帕斯(Scopus)

摘要

In this study, we developed a novel assay that simultaneously detects multiple miRNAs (microRNAs) within a single capillary by combining a tandem adenosine-tailed DNA bridge-assisted splinted ligation with denaturing capillary gel electrophoresis with laser-induced fluorescence. This proposed method not only represents a significant improvement in resolution but also allows for the detection of multiple miRNAs within a single capillary based on the length differences of specified target bridge DNA. The assay's linear range covers three orders of magnitude (1.0. nM to 1.0. pM) with a limit of detection (S/N. =3) as low as 190. fM (2.5. zmol). Five miRNAs of Epstein-Barr virus (EBV) were also detected in EBV-infected nasopharyngeal carcinoma cells, while they did not appear in non-virus infected cells. Moreover, the electropherogram indicated that the screening of isomiRs (isomer of miRNA) of BART2 by CE-LIF is feasible by our proposed method. The developed electrophoresis-based method for miRNA detection is fast, amplification-free, multiplexed and cost-effective, making it potentially applicable to large-scale screening of isomiRs.

原文英語
頁(從 - 到)2604-2610
頁數7
期刊Journal of Chromatography A
1218
發行號18
DOIs
出版狀態已出版 - 06 05 2011

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