Ribosome dynamics-based viability assay for Tobamovirus detection
A novel assay using ribosome dynamics to detect active Tobamoviruses by measuring translation viability, providing results in 8 hours. This approach combines Nanopore sequencing and qPCR to identify viable viruses, offering rapid and accurate diagnostics.
Overview
This innovative solution offers a rapid and precise method for detecting viable Tobamoviruses based on ribosome dynamics. By leveraging a molecular readout of virus viability, the assay focuses on identifying active virus genome translation, significantly accelerating detection compared to conventional RNA presence tests. The approach utilizes a combination of Nanopore sequencing and qPCR, delivering results within 8 hours, and provides a phenotypic test of virus viability.
Technical specifications
- Utilizes naturally occurring 5’-3’ RNA exonucleases for in vivo ribosome footprinting.
- Detects 5’ monophosphorylated mRNA degradation intermediates as indicators of active virus translation.
- Employs 5PSeq, a method that identifies characteristic ribosome stalling signatures.
- Simplified for field application with adapted oligos for RT of conserved Tobamovirus regions.
- Process includes seed germination, RNA extraction, ssRNA ligation, Tobamovirus RT and PCR, followed by Nanopore sequencing.
Technology readiness level
Currently at Technology Readiness Level 3, this technology has been validated in controlled laboratory settings. Future work includes developing a simplified diagnostic method, optimizing experimental protocols, and conducting proof-of-concept validation with over 100 samples to compare against existing diagnostics.
About 3N Bio AB
3N Bio AB is a Swedish life science startup and spin-off from SciLifeLab and Karolinska Institutet that develops diagnostic technologies to combat antimicrobial resistance. The company utilizes microbial metadegradomics to analyze RNA degradation signatures, enabling the direct, rapid measurement of bacterial growth and species-specific responses to antibiotics at the molecular level. By monitoring how bacterial cells recycle mRNAs, their approach functions without the need for complex experimental procedures, cell culture, or bacterial isolation, offering a streamlined method to detect antibiotic resistance within minutes.
The company aims to address the global threat of antibiotic-resistant bacteria and multidrug-resistant fungi by accelerating diagnosis and providing actionable information for antibiotic treatments. This technology is intended to support more personalized and rational antibiotic usage, potentially improving patient survival rates and containing the spread of resistant infections. Supported by research grants and scientific publications in journals like Nature Microbiology, 3N Bio is currently focused on applying its technology to develop simplified diagnostic kits for bacterial infections.
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