Real-time multiplexed PCR using surface enhanced Raman spectroscopy in a thermoplastic chip

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Molecular Diagnostics

Background:

• Surface-enhanced Raman spectroscopy (SERS) offers sensitive detection for various analytes.
• Current PCR-SERS assays require manual mixing, hindering real-time analysis.
• Multiplexed SERS detection following PCR is desirable for diagnostics.

Purpose of the Study:

• To develop a real-time PCR-SERS microsystem for simultaneous nucleic acid amplification and detection.
• To enable real-time, quantitative PCR with multiplexed SERS readout.
• To integrate PCR and SERS detection into a single, automated system.

Main Methods:

• A laser-cut thermoplastic fluidic chip with a dialysis membrane was designed.
• The chip separates the PCR reaction from a SERS-active silver colloid.
• Degradation of a Raman-reporter-labeled DNA probe releases the reporter for SERS detection.

Main Results:

• The microsystem achieved simultaneous nucleic acid amplification and real-time SERS detection.
• The system enabled real-time, single-well multiplexed readout of PCR products.
• Demonstrated simultaneous detection of two biomarker genes for MRSA.

Conclusions:

• The developed real-time PCR-SERS microsystem integrates amplification and detection.
• This approach eliminates manual steps and enables real-time monitoring.
• The system shows promise for rapid and multiplexed pathogen detection, such as for MRSA.