MutS protein-based fiber optic particle plasmon resonance biosensor for detecting single nucleotide polymorphisms

Area of Science:

• Biomedical Engineering
• Molecular Biology
• Nanotechnology

Background:

• Accurate and rapid detection of gene mutations is crucial for diagnosing genetic diseases and personalized medicine.
• Existing methods for mutation detection can be time-consuming, complex, or require extensive sample preparation.

Purpose of the Study:

• To develop a novel biosensing method for sensitive and rapid detection of gene mutations.
• To integrate the MutS protein with a fiber optic particle plasmon resonance (FOPPR) sensing system for enhanced detection capabilities.
• To validate the sensor's performance using clinical samples for potential point-of-care applications.

Main Methods:

• Conjugating MutS protein with gold nanoparticles (AuNPs) on an optical fiber core surface.
• Utilizing FOPPR to detect changes in light absorption caused by MutS binding to mismatched DNA.
• Employing a microfluidic chip for rapid sample analysis and real-time detection.

Main Results:

• Achieved a low limit of detection (0.49 nM) for AC mismatch heteroduplex DNA.
• Demonstrated a rapid analysis time of 15 minutes due to microfluidic integration.
• Successfully validated the method using a heterozygous PCR sample, determining an allelic fraction of 0.474.
• Showcased label-free, real-time determination of binding affinity and kinetics.

Conclusions:

• The MutS-functionalized FOPPR sensor offers a sensitive, rapid, and quantitative tool for detecting single nucleotide polymorphisms (SNPs).
• The system's ability to perform label-free, real-time analysis makes it suitable for various genetic applications.
• This technology holds promise for convenient point-of-care genetic testing and diagnostics.