On-site rpa-based lateral flow detection of plant genetic inserts, snps, and gmos

Overview

This solution offers a rapid, instrument-free genetic testing platform designed for real-world agricultural and environmental settings. By combining Recombinase Polymerase Amplification (RPA) with Lateral Flow Test (LFT) technology and using direct plant crude sap as a template, the platform enables on-site detection of plant genetic inserts, Single Nucleotide Polymorphisms (SNPs), and Genetically Modified Organisms (GMOs) at single-copy sensitivity. Results are available in approximately 25–30 minutes from sample collection, eliminating the need for laboratory infrastructure, nucleic acid purification, or trained technicians. The approach addresses a critical gap in field-based decision-making for crop management, disease outbreak response, food safety assurance, and environmental risk assessment, where traditional lab-based testing is too slow and impractical.

Technical specifications

• RPA-LFT assay amplifies target DNA at a single-copy detection limit without thermal cycling or specialized instruments
• Direct crude sap template eliminates the need for nucleic acid purification, reducing hands-on time and cost
• Alternative endonuclease enzyme replaces conventional Nfo-based chemistry, validated previously for detecting DNA and RNA viruses including CuLCrV, CYSDV, and RRV
• Detection sensitivity of up to 10 viral copies demonstrated in plant and insect samples in prior work
• Lateral flow readout provides clear visual results compatible with a small hand warmer for temperature control, enabling true field portability
• Multiplex capability under development to detect multiple targets simultaneously (PGI, SNPs, GMOs)
• Total assay time of approximately 25–30 minutes from sample collection to result

Technology readiness level

The underlying RPA-LFT methodology has been validated over more than a decade through the development of over fourteen on-site plant virus detection kits. The current research program aims to extend this proven platform to plant genetic inserts, SNPs, and GMOs through a phased plan: literature review, methodology development (3 months), assay validation including specificity, sensitivity, and environmental testing (4 months), field testing across diverse crops (3 months), and cost-effectiveness analysis (1.5 months). The approach is positioned for scalability and broad adoption by non-expert users in agricultural and environmental monitoring contexts.