期刊
ANALYTICAL CHEMISTRY
卷 94, 期 10, 页码 4311-4318出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.1c04988
关键词
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资金
- Ministry of Science and Technology (MOST) of Taiwan [MOST 1092221-E-007-006-MY3, MOST 110-2221-E-007-010-MY3]
- Taiwan's National Health Research Institutes [NHRI-EX110-11020EI]
- Ministry of Health and Welfare, Taiwan [MOHW110-TDU-B-211-144018]
Ovarian cancer is one of the most severe gynecologic cancers with asymptomatic early stages. This study developed an integrated microfluidic chip that can extract cfDNA from plasma, detect and quantify mutations in the OvCa biomarker BRCA1. By automating the cfDNA extraction and qPCR processes, this chip can be used for clinical screening of OvCa-associated mutations.
Ovarian cancer (OvCa) is among the most severe gynecologic cancers, yet individuals may be asymptomatic during its early stages. Routine, early screening for genetic abnormalities associated with OvCa could improve prognoses, and this can be achieved by detecting mutant genes in cell-free DNA (cfDNA). Herein, we developed an integrated microfluidic chip (IMC) that could extract cfDNA from plasma and automatically detect and quantify mutations in the OvCa biomarker BRCA1. The cfDNA extraction module relied on a vortex-type micromixer to mix cfDNA with magnetic beads surface-coated with cfDNA probes and could isolate 76% of molecules from a 200 mu L plasma sample in 45 min. The cfDNA quantification module, which comprised a micropump that evenly distributed 4.5 mu L of purified cfDNA into the on-chip, allele-specific quantitative polymerase chain reaction (qPCR) zones, was capable of quantifying mutant genes within 90 min. By automating the cfDNA extraction and qPCR processes, this IMC could be used for clinical screening for OvCa-associated mutations.
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