Viable tobamovirus seed test via peptide nucleic acid electrochemical sensor

Overview

The viable Tobamovirus seed test is an innovative solution that combines protease-based viability selection, rapid microfluidic PCR amplification, and sensitive electrochemical detection utilizing species-specific peptide nucleic acid (PNA) probes. This approach ensures RNA release only from capsid-intact, viable Tobamoviruses, thus preventing false positives from degraded or non-viable entities. Designed to accurately identify individual virus species, it aids in seed lot management and regulatory compliance, boosting confidence in seed exportation. The biosensor is highly specific, reusable for more than 10 cycles, cost-effective, and provides results within seven days. It is adaptable for other viruses and crops, with potential for automation and high-throughput testing.

Technical specifications

• Protease digestion selects for viable, capsid-protected virus particles.
• Microfluidic PCR allows for rapid and sensitive amplification of viral RNA.
• Species-specific PNA probes offer exceptional sequence specificity, reducing false positives.
• Gold nanoparticle-TiO₂ nanotube sensor ensures sensitive and repeatable electrochemical detection.
• Reusability: Sensor can be reused for 10 or more cycles.
• Adaptability: Suitable for various viruses and crops, ready for automation.
• Scalability: Cost-effective for high-throughput commercial lab testing.

Technology readiness level

This technology is currently at TRL 3, indicating experimental proof of concept. Future validation steps include optimizing protocols for selective RNA isolation, designing high-specificity PNA probes, and confirming the sensor's detection limits and repeatability. Further validation will assess the system's reusability, cost per test, and scalability for commercial applications. Partner support is sought for seed and virus standards, PCR data, lab access, and regulatory guidance.