Clinical Performance of Reverse Transcription Loop-mediated Isothermal Amplification COVID-19 Assay on Gold-nanoparticle-modified Screen-printed Carbon Electrode Using Differential Pulse Voltammetry

• Published in: Sensors and Materials
• Main Author: Zulhairee M.; Zain R.M.; Nor A.C.M.; Jafar N.F.; Noh M.F.M.; Noorden M.S.A.; Nordin A.N.; Rahim R.A.; Gunawan T.S.; Chin L.Y.; Mohd Y.; Azman N.A.; Zain Z.M.
• Format: Article
• Language: English
• Published: M Y U Scientific Publishing Division 2023
• Online Access: https://www.scopus.com/inward/record.uri?eid=2-s2.0-85176383035&doi=10.18494%2fSAM4430&partnerID=40&md5=a5a03085d2a02809c41ea8dcac02665a

The World Health Organization (WHO) has recommended real-time reverse transcription polymerase chain reaction (RT-PCR) as the gold standard for coronavirus disease detection. In this study, we aim to validate the clinical performance of reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay on a gold-nanoparticle-modified screen-printed carbon electrode (AuNP/SPCE) using differential pulse voltammetry (DPV) and to compare it with real-time RT-PCR. The electrodeposited AuNP on SPCE was quasi-spherical with a size of ±500 nm. The developed RT-LAMP primer was designed from the GenBank database using the NCBI Multiple Alignment tools and Jalview software. Nasopharyngeal clinical samples were obtained from suspected COVID-19 patients (n = 148). The RT-LAMP products were dropped on the modified AuNP/SPCE under DPV setting, which resulted in current change (∆I) responses. The positive and negative samples produced significantly different ∆I signals with a p-value <0.0001 at a 95% confidence interval using Student’s t-test. The RT-LAMP assay using Au/SPCE exhibited a 30 s response time per analysis. The clinical sensitivity and specificity obtained were 79.7 and 85.1%, respectively, with a detection limit of 0.4 copies µl−1. Hence, this proposed method is suitable for COVID-19 RNA detection in resource-limited settings. © 2023 M Y U Scientific Publishing Division. All rights reserved.