Rapid, Affordable, and Scalable SARS-CoV-2 Detection From Saliva

Area of Science:

• Molecular Biology
• Virology
• Diagnostics

Background:

• Accurate and rapid detection of SARS-CoV-2 is crucial for controlling the COVID-19 pandemic.
• Existing diagnostic methods can be time-consuming, expensive, or require specialized laboratory equipment.
• Point-of-care testing needs to be scalable, cost-effective, and easy to implement.

Purpose of the Study:

• To develop an inexpensive, rapid, and robust RT-LAMP assay for SARS-CoV-2 detection.
• To enable direct testing of patient saliva samples at point-of-care facilities and clinical labs.
• To achieve results within 30 minutes per reaction and less than 1 hour for 96 samples.

Main Methods:

• Utilized a novel combination of readily available reagents for RT-LAMP.
• Developed a method for bulk preparation, plating, and freezing of reagents for stability.
• Incorporated a dual reporter system (phenol red and SYTO-9) for internal validation and real-time monitoring.
• Adapted the assay for high-throughput automation in a 384-well format.

Main Results:

• Achieved direct detection of SARS-CoV-2 from patient saliva samples.
• Provided results in under 30 minutes per reaction, with total time under 1 hour for 96 samples.
• Demonstrated a cost of less than $2 per reaction.
• The dual reporter system ensured internal validation and assay reliability.

Conclusions:

• The developed RT-LAMP method is a scalable, cost-effective, and rapid diagnostic tool for SARS-CoV-2.
• This assay is suitable for point-of-care settings, improving accessibility to timely COVID-19 diagnosis.
• The method's robustness and ease of use facilitate high-throughput testing and automation.