Rhpcr lateral flow assay for rapid onsite GMO identification

Overview

This solution offers a rapid, portable testing platform for identifying genetically modified organisms (GMOs) in resource-limited and onsite settings. The workflow integrates a simplified heat lysis-based DNA isolation method, a miniaturized conventional PCR unit, RNase H-dependent PCR (rhPCR) primers for SNP-specific amplification, and lateral flow strips for visual detection of amplicons. Together, these components create a lab-in-a-suitcase-style system that can be operated by a minimally trained workforce, enabling decentralized GMO screening without the need for laboratory infrastructure or real-time PCR instrumentation. The approach is particularly valuable for regulatory agencies, seed companies, food processors, and inspection services requiring fast, field-deployable molecular diagnostics.

Technical specifications

Key features:

• Simple heat lysis-based DNA isolation compatible with crude extracts, eliminating the need for complex purification equipment
• Miniaturized conventional PCR unit enabling amplification outside laboratory environments
• rhPCR primers designed for SNP-specific gene target amplification, offering high specificity
• Primer labeling with FAM, biotin, and digoxigenin to facilitate lateral flow strip detection
• Dual-target lateral flow strips with an internal amplification control (IAC) for reliable result interpretation
• Inhibitor-resistant DNA polymerase formulations to handle crude sample matrices
• Validated workflow demonstrating 100% specificity in fish and shrimp SNP detection studies using 108 and 60 samples respectively

The assay combines the precision of SNP-based molecular detection with the simplicity of visual lateral flow readout, making it accessible for point-of-care and field-based applications.

Technology readiness level

The underlying workflow has been validated in two completed studies demonstrating 100% specificity, compatibility with crude DNA extracts, and reliable SNP detection using conventional PCR with lateral flow readout. Future validation activities include optimizing rapid DNA isolation protocols across multiple reagent candidates, evaluating inhibitor-resistant PCR master mixes for crude extract amplification, designing and optimizing rhPCR primers for GMO-specific gene or SNP targets, refining enzyme concentrations and PCR cycling conditions, and conducting full validation following AOAC Appendix J recommendations with target and non-target samples. The technology is currently at an advanced development stage with proven proof-of-concept data and a defined pathway toward standardized validation for GMO detection applications.