Construction of a novel peptide nucleic acid piezoelectric gene sensor microarray detection system

Area of Science:

• Biosensors
• Molecular Diagnostics
• Nanotechnology

Background:

• Conventional DNA probes exhibit limitations in specificity and efficiency for detecting nucleic acid sequences.
• Hepatitis B virus (HBV) genomic DNA detection is crucial for diagnosing and managing hepatitis B infections.

Purpose of the Study:

• To develop and optimize a novel piezoelectric gene sensor microarray for the direct detection of HBV genomic DNA.
• To evaluate the performance of bis-peptide nucleic acid (bis-PNA) probes compared to traditional DNA probes for HBV detection.

Main Methods:

• Construction of a 2x5 clamped-style piezoelectric gene sensor microarray with independently oscillating crystal units.
• Immobilization of bis-PNA probes onto the sensor surface for enhanced binding to target HBV DNA.
• Optimization of detection conditions, including buffer pH, ion concentration, and probe concentration.

Main Results:

• Optimal detection achieved with PBS buffer (pH 6.8), 20 mmol/L ion concentration, and 1.5 µmol/L probe concentration.
• Bis-PNA probes demonstrated significantly higher specificity for HBV DNA compared to DNA probes.
• A quantitative relationship was established between HBV DNA concentration (10 pg/L to 100 µg/L) and frequency decrease, with a linear regression equation (IgC = -2.7455 + 0.0691 ΔF) and high correlation coefficient (0.9923).

Conclusions:

• A novel piezoelectric gene sensor microarray with bis-PNA probes provides a sensitive and specific platform for HBV DNA detection.
• The optimized sensor system is suitable for quantitative detection of HBV in clinical samples, aligning with typical viral load ranges.