Rapid detection of seed-borne tobamoviruses using plant-dx 2.0 with CRISPR
PLANT-Dx 2.0 offers a rapid, field-deployable method for detecting seed-borne tobamoviruses using NASBA and CRISPR-Cas13a, capable of identifying viable viruses even in crude samples. This sensitive and accessible system is ideal for enhancing global seed trade safety.
Overview
PLANT-Dx 2.0 is an innovative solution for detecting seed-borne tobamoviruses, which pose significant threats to global agriculture. Utilizing nucleic acid sequence-based amplification (NASBA) and CRISPR-Cas13a, this system provides a rapid, sensitive, and field-deployable method for virus detection in seeds. Designed for flexibility, it can target a range of RNA and DNA viruses, including those with rod-shaped capsid structures like tobamoviruses. PLANT-Dx 2.0's ability to detect viral RNA at attomolar levels and operate with crude plant lysates makes it a valuable tool for agricultural biosecurity.
Technical specifications
Key features:
- NASBA and CRISPR-Cas13a integration: This combination allows for precise RNA virus detection.
- Programmable detection: Capable of targeting multiple virus types with computationally designed guide RNAs.
- Sensitivity levels: Offers a one-pot field version with femtomolar sensitivity and a two-step lab version with attomolar sensitivity.
- Versatile operation: Can be used with minimal equipment, including handheld light sources or plate-based readers for high-throughput analysis.
- Platinum chloride viability assay: Enhances detection specificity by differentiating between viable and non-viable virus particles.
Technology readiness level
Currently at TRL 4, PLANT-Dx 2.0 has been validated in controlled laboratory settings. Future development aims include optimizing the assay for seed testing, enhancing detection sensitivity, and establishing practical limits for detecting viable tobamoviruses in seed samples. This will involve spiking seed lysates with known virus quantities and varying healthy seed mixtures to assess effectiveness and reliability.
About Stemloop, Inc.
Stemloop, Inc. was a biotechnology company founded in 2019 that specialized in cell-free synthetic biology to create programmable, paper-based biosensors. Developed from academic research at Northwestern University, the company’s platform utilized DNA, RNA, and protein components to detect specific chemical analytes and contaminants in environmental and health contexts. The technology was designed to function without the need for extensive laboratory infrastructure, instead providing rapid, inexpensive, and on-site testing capabilities through simple visual signals, such as color changes, when a target substance was present. The company originated in Evanston, Illinois, and received support from various research programs, including the National Science Foundation and the National Institute of Environmental Health Sciences.
The company focused on democratizing access to testing for critical threats, with a flagship application being the µSense lead-in-water test designed to improve household safety and water quality monitoring. By decentralizing diagnostic capabilities, Stemloop aimed to address challenges that previously required costly and time-consuming analytical chemistry techniques. The company sought to apply its biosensing technology across multiple sectors, including environmental monitoring, agriculture, biomanufacturing, and human health, to deliver actionable information for risk mitigation. Stemloop, Inc. concluded its operations in 2025.
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