Fabrication of Cost-Effective Microchip-Based Device Using Sandblasting Technique for Real-Time Multiplex PCR...

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Microfluidics

Background:

• Multiplex PCR (mPCR) and microfluidic technologies are vital for biomedical applications.
• Current microfluidic diagnostics struggle with multi-target detection due to limited channels, complex fabrication, and high costs.

Purpose of the Study:

• To develop a cost-effective sandblasting method for manufacturing silicon microchips for field mPCR detection.
• To overcome biocompatibility issues and optimize the microchip surface for enhanced molecular detection.

Main Methods:

• Fabrication of silicon microchips using a sandblasting method.
• Surface modification with bovine serum albumin (BSA) to improve biocompatibility.
• Optimization of BSA coating conditions using SEM and EDS.
• On-chip PCR and mPCR assays for COVID-19 and resistance genes.

Main Results:

• Sandblasted microchips showed a rough surface, requiring BSA coating for improved biocompatibility.
• Optimized BSA coating (65 °C for 60 min) enhanced performance.
• Achieved rapid on-chip PCR for COVID-19 (500 copies/mL in 20 min) with higher amplification than dry-etched chips.
• Successfully performed 50-minute mPCR for five resistance genes with high specificity and reliability.

Conclusions:

• The sandblasted microchip platform offers a rapid, accessible, and cost-effective solution for multiplex molecular detection.
• This method enables large-scale microfabrication conveniently and affordably.
• The developed microdevice is suitable for field-based mPCR screening of pathogens and resistance genes.