Gelred® and gelgreen® based detection method for tobamoviruses
A novel detection method utilizing GelRed® and GelGreen® for the safe and non-destructive identification of Tobamoviruses in seeds using RT-PCR. This method is environmentally safe and discriminates between viable and non-viable viruses.
Overview
This innovative solution leverages GelRed® (GR) and GelGreen® (GG), advanced nucleic acid staining reagents, for the detection of Tobamoviruses in seed samples. These reagents provide a safe and environmentally friendly approach to virus detection, as they only bind to naked DNA or RNA, ensuring they do not penetrate living cells. The method is structured in two phases: a non-destructive phase for isolating non-viable nucleic acids and a destructive phase for detecting total nucleic acids. This approach is particularly useful for accurately distinguishing between viable and non-viable Tobamoviruses using RT-PCR.
Technical specifications
- Nucleic Acid Binding: GR and GG bind specifically to naked DNA or RNA, enabling non-destructive analysis of seed samples.
- Two-Phase Methodology:
- Non-destructive phase: Isolates non-viable nucleic acids using PBS and detergent/proteinase K, followed by spectrophotometric quantification and RT-PCR.
- Destructive phase: Isolates total RNA and dsRNA for comprehensive RT-PCR analysis.
- RT-PCR Testing: Utilizes primers for conserved regions of the Tobamovirus genome, including the replicase gene and the coat protein gene.
- Environmentally Safe: GR and GG are non-hazardous and do not penetrate living cells, making the method safe for both the environment and users.
Technology readiness level
Currently at TRL 2, this method is in the early stages of development and experimentation. Planned validation includes virus propagation and maintenance, seed collection, method standardization, and eventual validation from greenhouse and field samples over an 18-month period.
About Brock University
Brock University is a comprehensive public institution serving the Niagara region of Ontario, combining research, experiential education, and community engagement. Industry partners access co‑located collaboration space and prototyping resources in the campus innovation hub, while a mature co‑op program connects R&D teams with job‑ready talent. Its location near the U.S. border places companies within the cross‑border Great Lakes manufacturing and logistics corridor and alongside the region’s established grape‑and‑wine ecosystem. Research is supported by competitive funding from Canada’s Tri‑Agency councils and provincial programs, alongside industry partnerships. A dedicated technology transfer office guides IP strategy, licensing, and startup formation, providing clear paths from sponsored research to market.
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