INSIGHT: A population-scale COVID-19 testing strategy combining point-of-care diagnosis with centralized...

Area of Science:

• Molecular Biology
• Infectious Disease Diagnostics
• Genomics

Background:

• Accurate and scalable diagnostic tools are crucial for managing infectious disease outbreaks like coronavirus disease 2019.
• Current testing methods may face limitations in speed, scalability, or decentralized application.

Purpose of the Study:

• To develop and present INSIGHT (isothermal NASBA sequencing-based high-throughput test), a novel two-stage testing strategy for coronavirus disease 2019.
• To combine point-of-care diagnosis with next-generation sequencing for population-level testing capabilities.

Main Methods:

• Utilized a barcoded isothermal NASBA (nucleic acid sequence-based amplification) reaction in two stages.
• Stage 1: Rapid, decentralized readout (1-2 hours) with fluorescence or lateral flow, incorporating sample-specific barcodes.
• Stage 2: Pooled reaction products from Stage 1 undergo highly multiplexed, sequencing-based confirmation and centralized data collection.

Main Results:

• Achieved a 95% limit of detection below 50 copies of viral RNA per reaction.
• Demonstrated a two-stage approach combining rapid point-of-care testing with scalable, high-throughput sequencing.
• INSIGHT facilitates early isolation of presymptomatic/asymptomatic patients and centralized data management.

Conclusions:

• INSIGHT offers a promising strategy for population-scale coronavirus disease 2019 testing.
• The system is suitable for development into rapid home-based or point-of-care assays.
• The methodology holds potential for scalable infectious disease surveillance.