Pcr-based assay for detecting viable tobamoviruses on seeds
A novel PCR-based assay using PGM-MBs and RNase treatment with RT-qPCR for sensitive detection of viable tobamoviruses on seeds, enhancing virus detection sensitivity by up to 64 times and ensuring intact viral detection.
Overview
This innovative solution introduces a novel PCR-based assay designed specifically for the sensitive detection of viable tobamoviruses on seeds. By leveraging porcine gastric mucin-conjugated magnetic beads (PGM-MBs) and RNase treatment followed by a two-step RT-qPCR process, this method significantly enhances the sensitivity and specificity of virus detection. The PGM-MBs facilitate the concentration of virus particles from seed washing solutions, while the RNase treatment ensures that only viruses with intact capsids and genomes are detected. This technology promises to improve the accuracy of detecting infectious tobamoviruses, crucial for agricultural biosecurity.
Technical specifications
Key features:
- PGM-MBs: These beads absorb biomolecules from virus surfaces, allowing magnetic concentration of virus particles.
- RNase Treatment: Degrades viral genomes with damaged capsids, ensuring only viable viruses are detected.
- Two-Step RT-qPCR: Detects the entire viral genome by reverse transcribing from the 3' end and quantifying cDNA from the 5' end.
Applications:
- Detection of tobamoviruses in agricultural seeds, enhancing crop safety and quality.
- Suitable for diverse virus types, thanks to customizable primers for genetically varied strains.
Technology readiness level
This technology is at TRL 6, indicating a prototype demonstrated in a relevant environment. It has been validated in municipal wastewater settings and is now being optimized for seed washing solutions. Future validation will include testing on tomato seeds to determine detection limits and optimize assay performance.
About University of Florida
The University of Florida is a comprehensive public research university with a broad academic and research portfolio and a statewide presence. Industry collaborates through co-located labs and shared core facilities and through an integrated academic health system that accelerates clinical translation. A statewide extension network and multiple research and education sites connect companies with field-scale testing and rapid deployment, while incubators and an adjacent innovation district provide pathways from lab to market. Research is supported by competitive funding from major federal agencies such as NIH, NSF, USDA, and DOE. A dedicated technology transfer office supports IP, licensing, and startup formation.
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