On-chip quantitative detection of pathogen genes by autonomous microfluidic PCR platform

Area of Science:

• Biotechnology
• Microfluidics
• Molecular Diagnostics

Background:

• Polymerase Chain Reaction (PCR) is vital for clinical and food testing.
• Need for rapid, portable, and affordable PCR devices for on-site applications.

Purpose of the Study:

• Develop an autonomous, disposable microfluidic PCR chip for quantitative detection of pathogens.
• Enable rapid, on-site genetic analysis with minimal user intervention.

Main Methods:

• Designed a novel microfluidic chip using injection-molded cyclo-olefin polymer (COP).
• Incorporated a unique driving microchannel to control capillary flow for autonomous operation.
• Utilized non-ionic surfactant coating for optimal hydrophilic surface properties.

Main Results:

• Achieved quantitative real-time PCR detection of human, E. coli, and E. coli O157 DNA.
• Demonstrated successful detection with low genomic DNA concentrations (down to 0.0019 pg/μl for E. coli).
• Completed PCR analysis in under 18 minutes.

Conclusions:

• The developed microfluidic PCR chip offers a rapid, user-friendly, and low-cost solution.
• Potential for widespread use in on-site gene testing applications.
• Advances portable molecular diagnostics for field use.