Membrane-Based In-Gel Loop-Mediated Isothermal Amplification (mgLAMP) System for SARS-CoV-2 Quantification in...

Area of Science:

• Environmental science
• Molecular biology
• Biotechnology

Background:

• The COVID-19 pandemic's transition to endemic status necessitates robust environmental surveillance for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2).
• Accurate quantification of SARS-CoV-2 in ambient waters is crucial for early outbreak detection and public health monitoring.

Purpose of the Study:

• To develop a rapid, point-of-use system for quantifying SARS-CoV-2 in environmental water samples.
• To create a user-friendly method suitable for large-scale environmental surveillance without specialized equipment or extensive training.

Main Methods:

• Development of a membrane-based in-gel loop-mediated isothermal amplification (mgLAMP) system.
• Integration of viral concentration, in-assay lysis, and hydrogel-based RT-LAMP quantification with enhanced fluorescence detection.
• Utilization of a 3D-printed portable device for incubation and fluorescence illumination, coupled with smartphone imaging and machine learning for result interpretation.

Main Results:

• The mgLAMP system achieved a sample-to-result time of under 1 hour.
• Detection of SARS-CoV-2 below 0.96 copies/mL in Milli-Q water.
• In surface water, mgLAMP detected down to 93 copies/mL, outperforming RT-qPCR which was inhibited at 930 copies/mL.

Conclusions:

• The mgLAMP system demonstrates high sensitivity and efficiency for SARS-CoV-2 quantification in environmental waters.
• This method is a promising tool for accessible, large-scale environmental surveillance of SARS-CoV-2.
• The portable, user-friendly design facilitates widespread adoption in public health monitoring efforts.