Viability dpcr for identifying infectious tobamovirus in seeds
A novel method combining Promega Viability PCR reagent and digital PCR to detect viable tobamoviruses in seeds, offering 1000x sensitivity over ELISA and distinguishing infectious from non-infectious viruses in under 24 hours.
Overview
This innovative approach harnesses the power of the Promega Viability PCR reagent (PVPR) in conjunction with digital PCR (dPCR) to accurately determine the presence of viable tobamovirus in seeds. The method distinguishes between viable and non-viable viruses, providing results in less than 24 hours and offering sensitivity that is approximately 1000 times greater than traditional ELISA methods. This solution reduces the number of seeds required for testing and enhances the accuracy of viral detection, making it a valuable tool for the agricultural industry.
Technical specifications
The PVPR system binds covalently to the genetic material of viruses with compromised capsids (non-viable viruses), preventing their genomic material from being amplified by PCR. This allows for the selective amplification of only intact, viable tobamoviruses during the dPCR step. The method involves two multiplexed dPCR reactions covering the tobamovirus genome, confirming the presence of complete, viable genomes. The solution is adaptable to multiple viruses or bacteria, further enhancing its applicability across various sample types.
Technology readiness level
Currently at a Technology Readiness Level 2, this method is in the early stages of validation. The initial steps include confirming the PVPR's effectiveness in binding to tobamovirus RNA with damaged capsids using a quantified ATCC tobamovirus sample, and designing a comprehensive dPCR assay to assess viral genome presence. Further validation will confirm the method's capability to accurately quantify viable tobamoviruses.
About Michigan Technological University
Michigan Technological University is a mid‑sized public technological research university known for hands‑on, industry‑engaged innovation and a strong applied research culture. Companies tap co‑op pathways and an embedded, multi‑year enterprise model to access talent while shaping solutions alongside faculty and students. Co‑located laboratories, pilot‑scale facilities, and instrumented field sites—including cold‑weather test ranges near campus, waterfront facilities on the Great Lakes, and a research presence in Southeast Michigan—enable validation from lab to operational environments. Research is supported by competitive awards from agencies such as the National Science Foundation, the Department of Energy, the Department of Defense, and NASA, alongside state and industry sponsors. A dedicated technology transfer office streamlines IP protection, sponsored research agreements, and startup formation for corporate collaborators.
delivered to your inbox.