Integrated Microfluidic System for Cell-Free DNA Extraction from Plasma for Mutant Gene Detection and Quantification

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Gynecologic Oncology

Background:

• Ovarian cancer (OvCa) is a severe gynecologic malignancy often diagnosed late due to asymptomatic early stages.
• Early detection through genetic screening of cell-free DNA (cfDNA) can significantly improve patient prognoses.
• Identifying specific mutations, such as in the BRCA1 gene, is key for targeted OvCa screening.

Purpose of the Study:

• To develop an integrated microfluidic chip (IMC) for automated cfDNA extraction and mutation detection.
• To enable rapid and efficient quantification of OvCa-associated mutations, specifically in the BRCA1 gene.
• To assess the potential of the IMC for clinical application in early ovarian cancer screening.

Main Methods:

• Development of an IMC featuring a cfDNA extraction module utilizing magnetic beads and a vortex-type micromixer.
• Integration of a cfDNA quantification module with a micropump for on-chip, allele-specific quantitative polymerase chain reaction (qPCR).
• Automated processing of plasma samples for cfDNA isolation and subsequent mutation analysis.

Main Results:

• The cfDNA extraction module achieved 76% molecule isolation from 200 μL plasma within 45 minutes.
• The cfDNA quantification module completed mutation analysis within 90 minutes.
• The automated IMC demonstrated capability for detecting and quantifying BRCA1 mutations in cfDNA.

Conclusions:

• The developed IMC successfully automates cfDNA extraction and qPCR, streamlining mutation detection.
• This integrated system offers a promising tool for the clinical screening of ovarian cancer-associated mutations.
• Automated analysis of cfDNA mutations via microfluidics holds potential for improving early diagnosis and management of OvCa.