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J Environ Chem Eng . Loop-mediated isothermal amplification-based electrochemical sensor for detecting SARS-CoV-2 in wastewater samples

tetano

Editor, Senior Moderator
J Environ Chem Eng


. 2022 Jun;10(3):107488.
doi: 10.1016/j.jece.2022.107488. Epub 2022 Feb 28.
Loop-mediated isothermal amplification-based electrochemical sensor for detecting SARS-CoV-2 in wastewater samples


Roberto G Ramírez-Chavarría[SUP] 1 [/SUP], Elizabeth Castillo-Villanueva[SUP] 1 2 [/SUP], Bryan E Alvarez-Serna[SUP] 1 [/SUP], Julián Carrillo-Reyes[SUP] 3 [/SUP], Rosa María Ramírez-Zamora[SUP] 1 [/SUP], Germán Buitrón[SUP] 3 [/SUP], Luis Alvarez-Icaza[SUP] 1 [/SUP]



Affiliations

Abstract

The current pandemic COVID-19 caused by the coronavirus SARS-CoV-2, has generated different economic, social and public health problems. Moreover, wastewater-based epidemiology could be a predictor of the virus rate of spread to alert on new outbreaks. To assist in epidemiological surveillance, this work introduces a simple, low-cost and affordable electrochemical sensor to specifically detect N and ORF1ab genes of the SARS-CoV-2 genome. The proposed sensor works based on screen-printed electrodes acting as a disposable test strip, where the reverse transcription loop-mediated isothermal amplification (RT-LAMP) reaction takes place. Electrochemical detection relies upon methylene blue as a redox intercalator probe, to provide a diffusion-controlled current encoding the presence and concentration of RT-LAMP products, namely amplicons or double-stranded DNA. We test the performance of the sensor by testing real wastewater samples using end-point and time course measurements. Results show the ability of the electrochemical test strip to specifically detect and quantify RT-LAMP amplicons below to ~ 2.5 × 10[SUP]-6[/SUP] ng/μL exhibiting high reproducibility. In this sense, our RT-LAMP electrochemical sensor is an attractive, efficient and powerful tool for rapid and reliable wastewater-based epidemiology studies.

Keywords: Electrochemical sensor; Epidemiology; RT-LAMP; Redox intercalator; SARS-CoV-2; Wastewater.
 
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