Rapid real-time recirculating PCR using localized surface plasmon resonance (LSPR) and piezo-electric pumping

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Microfluidics

Background:

• Current methods for bloodstream infection (BSI) pathogen detection are often slow or lack comprehensive diagnostic data.
• Real-time polymerase chain reaction (qPCR) offers specific detection and quantification but requires technological advancements for rapid, multiplex clinical use.

Purpose of the Study:

• To demonstrate the feasibility of a novel microfluidic platform for rapid, multiplex qPCR analysis.
• To integrate localized surface plasmon resonance (LSPR) imaging with qPCR for label-free DNA hybridization detection.

Main Methods:

• Development of a microfluidic chip with a recirculating design for real-time analysis.
• Integration of highly sensitive, label-free LSPR imaging to monitor DNA hybridization.
• Demonstration of single-target and multiplex DNA amplification detection.

Main Results:

• Achieved a limit of detection of 5 fg μL^-1 for E. coli DNA in single-target PCR, equivalent to approximately 300 bacteria/mL.
• Successfully demonstrated simultaneous real-time detection of multiple target genes.
• Platform performance achieved within a 15-minute timeframe.

Conclusions:

• The novel microfluidic platform shows significant potential for rapid, multiplex pathogen detection in clinical settings.
• This technology could offer life-saving benefits for patients with bloodstream infections by enabling faster diagnosis and treatment planning.