Loop-mediated isothermal amplification-based electrochemical sensor for detecting SARS-CoV-2 in wastewater samples

Area of Science:

• Environmental Science
• Biotechnology
• Public Health

Background:

• The COVID-19 pandemic, caused by SARS-CoV-2, presents significant global health and economic challenges.
• Wastewater-based epidemiology serves as a crucial early warning system for tracking virus spread and potential outbreaks.

Purpose of the Study:

• To develop a simple, affordable electrochemical sensor for specific detection of SARS-CoV-2 genes (N and ORF1ab).
• To enable rapid and reliable wastewater-based epidemiological surveillance.

Main Methods:

• Utilized screen-printed electrodes as disposable test strips for reverse transcription loop-mediated isothermal amplification (RT-LAMP).
• Employed methylene blue as a redox intercalator probe for electrochemical detection of RT-LAMP amplicons (double-stranded DNA).
• Validated sensor performance using real wastewater samples with end-point and time-course measurements.

Main Results:

• The electrochemical sensor demonstrated specific detection and quantification of SARS-CoV-2 RT-LAMP amplicons.
• Achieved high reproducibility with detection limits below 2.5 × 10^-6 ng/μL.
• Successfully tested on real wastewater samples, confirming its practical applicability.

Conclusions:

• The developed RT-LAMP electrochemical sensor is an effective tool for wastewater-based epidemiology.
• Offers a rapid, reliable, and accessible method for SARS-CoV-2 surveillance in environmental samples.
• Supports public health strategies through timely detection of viral spread.