Development of an Electrochemical Sensor for SARS-CoV-2 Detection Based on Loop-Mediated Isothermal Amplification

• Published in: Biosensors (Basel) Vol. 13; no. 10; p. 924
• Main Authors: Rivas-Macho, Ane, Eletxigerra, Unai, Diez-Ahedo, Ruth, Barros, Ángela, Merino, Santos, Goñi-de-Cerio, Felipe, Olabarria, Garbiñe
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 01.10.2023; MDPI
• Subjects: Adhesives; Antigens; biosensor; Biosensors; Chemical sensors; Coronaviruses; COVID-19; Deoxyribonucleic acid; Design and construction; DNA; Electrochemical analysis; Electrochemical apparatus; electrochemical detection; Electrochemistry; Electrodes; Gene amplification; Inactivation; Laboratory equipment; loop-mediated isothermal amplification (LAMP); Methods; Methylene blue; Pandemics; SARS-CoV-2; Sensors; Severe acute respiratory syndrome coronavirus 2; Viral diseases; Viruses; Spain; United Kingdom > UK; Germany
• ISSN: 2079-6374; 2079-6374
• DOI: 10.3390/bios13100924

The pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused more than 6 million deaths all over the world, demonstrating the need for a simple, fast and cost-effective point-of-care (POC) test for the detection of the virus. In this work, we developed an electrochemical sensor for SARS-CoV-2 virus detection on clinical samples based on loop-mediated isothermal amplification (LAMP). With the development of this novel sensor, the time of each measurement is significantly reduced by avoiding the DNA extraction step and replacing it with inactivation of the sample by heating it at 95 °C for 10 min. To make the reaction compatible with the sample pre-treatment, an RNase inhibitor was added directly to the premix. The LAMP product was measured in a novel, easy-to-use manufactured sensor containing a custom-made screen-printed carbon electrode. Electrochemical detection was performed with a portable potentiostat, and methylene blue was used as the redox-transducing molecule. The developed sensor achieved a limit of detection of 62 viral copies and was 100% specific for the detection of the SARS-CoV-2 virus. The performance of the electrochemical sensor was validated with nasopharyngeal samples, obtaining a sensibility and specificity of 100% compared to the gold standard RT-PCR method.