Rt-qpcr with DMS footprinting for viable tobamovirus detection
A novel RT-qPCR method using DMS footprinting to distinguish viable Tobamovirus from damaged particles, enhancing viral detection accuracy and compatibility with existing workflows.
Overview
This innovative approach leverages RT-qPCR with dimethyl sulfate (DMS) footprinting to differentiate viable Tobamovirus particles from damaged ones. The method utilizes the selective modification of exposed RNA in non-viable virions, ensuring only viable material is detected. This enhances accuracy in viral detection and is compatible with existing PCR workflows.
Technical specifications
- Selective RNA modification: DMS and other alkylating agents selectively modify RNA in damaged or non-viable virions, blocking reverse transcription and PCR amplification.
- Compatibility: The approach is analogous to PMS treatment used in microbial detection, ensuring seamless integration with established laboratory workflows.
- Patented MSR-seq technique: Used to map methylation patterns, optimizing sample treatment and qPCR assays for dynamic range determination between viable and damaged virions.
- Targeted qPCR development: Focuses on labile regions of the RNA genome that become exposed due to disinfection, ensuring precise detection of viable virions.
Technology readiness level
This technology is at TRL 3, indicating that the method has been formulated and is in the experimental proof-of-concept stage, with ongoing validation required using real-world samples from partners.
About MesoRNA
MesoRNA specializes in advanced sequencing technologies for small non-coding RNAs, known as mesoRNAs, which range from 30 to 300 nucleotides. The company has developed its proprietary MSR-seq technology to enable comprehensive, multiplexed next-generation sequencing (NGS) library preparation. This platform allows researchers to measure tens of thousands of unique molecules per sample with high base-calling accuracy. Additionally, the company provides a full-service laboratory option for sample processing and is developing a TL-qPCR assay intended to translate biomarker discoveries into diagnostic applications. By addressing the technical challenges historically associated with studying these pervasive but under-researched molecules, MesoRNA aims to complete the understanding of the central dogma of molecular biology.
These tools serve academic laboratories, core facilities, and diagnostic developers who require precise, scalable methods to analyze the biological roles of mesoRNAs. By streamlining the workflow from library preparation to bioinformatics analysis, the company helps users overcome previous limitations in small RNA research. MesoRNA, founded in 2023 and based in Chicago, grew out of over a decade of academic research led by Dr. Tao Pan. The company provides a crucial resource for institutions seeking to investigate the regulatory functions of non-coding RNAs, which constitute a significant majority of human cellular nucleic acids, potentially unlocking new pathways for biological discovery and diagnostic development.
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