Highly multiplex nucleic acid assay for pathogen-free solanaceous crops

Overview

Fluid Discovery introduces an innovative nucleic acid assay designed to ensure pathogen-free solanaceous crops, specifically targeting tobamovirus infections. This highly multiplexed solution combines droplet digital PCR (ddPCR) with capillary electrophoresis (CE) for enhanced detection and discrimination of viral species and strains. Utilizing viability-selective pretreatments like propidium monoazide (PMA) and ethidium monoazide (EMA), the assay distinguishes between viable and non-viable pathogens efficiently. With the ability to measure over 100 targets in a single assay, it offers a consolidated approach to seed health monitoring, reducing the risk of false-negatives due to viral genetic diversity.

Technical specifications

• Multiplex Capability: Measures over 100 nucleic acid targets per reaction, allowing comprehensive viral monitoring.
• Viability Discrimination: Utilizes PMA/EMA or RNase pretreatments to differentiate viable from non-viable virus particles.
• Automation and Speed: Fully automatable on standard laboratory equipment, with a complete assay time of under 7 hours.
• Targeted Pathogens: Focuses on tobamoviruses affecting solanaceous crops, including TMV, ToMV, ToMMV, ToBRFV, TMGMV, PMMoV, and PaMMV.
• Proprietary Primer Design: Uses a proprietary primer pipeline to ensure specificity and redundancy, optimizing detection across viral genetic diversity.

Technology readiness level

Currently at TRL 4, this technology has undergone laboratory validation with various protocols and primer designs. Future validation includes expanding the panel and confirming the assay's effectiveness on real seed samples using automated protocols, ensuring readiness for broader application.