Centrifugal microfluidic system for colorimetric sample-to-answer detection of viral pathogens

Area of Science:

• Food Safety
• Microfluidics
• Molecular Diagnostics

Background:

• Foodborne viral pathogens pose a significant public health risk.
• Current detection methods can be time-consuming and complex.
• Need for rapid, automated diagnostic tools for food safety.

Purpose of the Study:

• To develop and validate an automated microfluidic system for multiplexed detection of foodborne viruses.
• To achieve a sample-to-answer format for efficient pathogen identification.
• To enhance food safety surveillance and outbreak response capabilities.

Main Methods:

• A centrifugal microfluidic platform automating 24 assay steps, including thermal lysis, PCR, hybridization, and colorimetric detection.
• Fabrication of microfluidic cartridges from thermoplastic polymers with integrated micropillar arrays.
• Development of oligonucleotide primers and probes for multiplexed identification of five viruses, including norovirus and hepatitis A virus (HAV).
• Utilized digoxigenin-labeled amplicons and immunoenzymatic conversion for colorimetric readout.

Main Results:

• Accurate and repetitive discrimination of positive and negative HAV signals at 350 genome copies/μL demonstrated using endpoint detection.
• Optimization studies revealed potential signal variations due to micropillar array configurations.
• Numerical modeling supported findings on flow behavior and signal distribution.

Conclusions:

• The automated microfluidic system offers a rapid, sample-to-answer solution for detecting foodborne viral pathogens.
• This technology has the potential to significantly reduce response times in food safety monitoring and outbreak investigations.
• Automation is key to advancing rapid pathogen detection for public health protection.