Isolation and Quantification of Methylated Cell-Free DNA in Plasma on an Integrated Microfluidic System

Yu Jen Cheng, Chih Hung Wang, Keng Fu Hsu, Gwo Bin Lee

研究成果: Article同行評審

1 引文 斯高帕斯(Scopus)

摘要

Methylated cell-free DNA (cfDNA) has been deemed a promising biomarker for ovarian cancer (OvCa) prognosis and therapy selection. However, exploring the methylation profiles of tumor suppressor genes in cfDNA remains a challenge due to their extremely low concentrations and complicated protocols, as well as methodological constraints. In this study, an integrated microfluidic system was developed to automatically (1) capture methylated cfDNA in plasma by magnetic beads coated with the methyl-CpG-binding domain and (2) quantify the methylation level of tumor suppressor genes by on-chip quantitative polymerase chain reaction (qPCR). For capturing methylated cfDNA from a very small amount of plasma, samples along with beads were mixed in a new micromixer to enhance the capture rate. With a high capture rate (72%) and a limit of quantification of 0.1 pg/μL (3 orders of magnitude lower than that of the benchtop method), the compact system could detect the methylated cfDNA from only 20 μL of plasma sample in 2 h. Furthermore, the dynamic range, from 0.1 to 2000 pg/μL of methylated cfDNA, spans the physiological range in plasma, signifying that this device has great potential for personalized medicine in OvCa.

原文English
頁(從 - 到)2134-2141
頁數8
期刊Analytical Chemistry
94
發行號4
DOIs
出版狀態Published - 2022 2月 1

All Science Journal Classification (ASJC) codes

  • 分析化學

指紋

深入研究「Isolation and Quantification of Methylated Cell-Free DNA in Plasma on an Integrated Microfluidic System」主題。共同形成了獨特的指紋。

引用此