期刊
ANALYTICAL CHEMISTRY
卷 94, 期 4, 页码 2134-2141出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.1c04471
关键词
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资金
- Ministry of Science and Technology (MOST) of Taiwan [MOST 109-2221-E007-006-MY3]
- Taiwan's National Health Research Institutes [NHRI-EX110-11020EI, NHRI110A1-CACO-02212111]
In this study, an integrated microfluidic system was developed to automatically capture and quantify the methylation level of cfDNA. The system achieved high capture rate and low limit of quantification, allowing for efficient detection of methylated cfDNA from plasma samples in a short time.
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/mu L (3 orders of magnitude lower than that of the benchtop method), the compact system could detect the methylated cfDNA from only 20 mu L of plasma sample in 2 h. Furthermore, the dynamic range, from 0.1 to 2000 pg/mu L of methylated cfDNA, spans the physiological range in plasma, signifying that this device has great potential for personalized medicine in OvCa.
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