Fluorescent paper-based DNA sensor using pyrrolidinyl peptide nucleic acids for hepatitis C virus detection

Area of Science:

• Biomedical Engineering
• Molecular Diagnostics
• Materials Science

Background:

• Hepatitis C virus (HCV) infection poses a significant global health challenge.
• Accurate and accessible diagnostic tools are crucial for effective HCV management.
• Current diagnostic methods may be limited by cost, complexity, or accessibility.

Purpose of the Study:

• To develop a novel, sensitive, and selective fluorescent paper-based DNA sensor for HCV detection.
• To utilize a specific pyrrolidinyl peptide nucleic acid (acpcPNA) probe for enhanced detection accuracy.
• To create a low-cost, portable system suitable for point-of-care applications.

Main Methods:

• Covalent immobilization of acpcPNA probes onto cellulose paper via reductive alkylation.
• Fluorescence-based detection using a fluorescent dye and a portable camera gadget with a smartphone.
• Optimization of reaction conditions for sensitive and selective DNA detection.

Main Results:

• A linear relationship was observed between fluorescence change and HCV DNA concentration (5-100 pmol) with a high correlation coefficient (0.9956).
• The sensor achieved a low limit of detection (5 pmol) for synthetic oligonucleotides.
• The acpcPNA probe demonstrated high selectivity against mismatched and non-complementary DNA targets.
• Successful detection of HCV complementary DNA (cDNA) from clinical samples was achieved, indicating real-world applicability.

Conclusions:

• The developed fluorescent paper-based sensor offers a sensitive, selective, and label-free method for DNA detection.
• The acpcPNA probe provides high specificity, crucial for accurate viral detection.
• This technology holds significant promise for low-cost, rapid, and accessible point-of-care diagnostics for HCV.