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S&M3426 Research Paper of Special Issue https://doi.org/10.18494/SAM4430 Published: October 25, 2023 Clinical Performance of Reverse Transcription Loop Mediated Isothermal Amplification COVID-19 Assay on Gold- nanoparticle-modified Screen-printed Carbon Electrode Using Differential Pulse Voltammetry [PDF] Munirah Zulhairee, Rozainanee Mohd Zain, Adibah Che Mohamad Nor, Nur Fadhilah Jafar, Mohd Fairulnizal Md Noh, Mohd Shihabuddin Ahmad Noorden, Anis Nurashikin Nordin, Rosminazuin Ab Rahim, Teddy Surya Gunawan, Lim Ying Chin, Yusairie Mohd, Nur Asyiqin Azman, and Zainiharyati Mohd Zain (Received April 17, 2023; Accepted July 24, 2023) Keywords: COVID-19, RT-PCR, RT-LAMP, AuNP/SPCE, differential pulse voltammetry, electrochemical sensor
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 gold-nanoparticle-modified screen-printed carbon electrode (AuNP/SPCE) using differential pulse voltammetry (DPV) and to compare it with real-time RT-PCR. The shape of 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.
Corresponding author: Zainiharyati Mohd ZainThis work is licensed under a Creative Commons Attribution 4.0 International License. Cite this article Munirah Zulhairee, Rozainanee Mohd Zain, Adibah Che Mohamad Nor, Nur Fadhilah Jafar, Mohd Fairulnizal Md Noh, Mohd Shihabuddin Ahmad Noorden, Anis Nurashikin Nordin, Rosminazuin Ab Rahim, Teddy Surya Gunawan, Lim Ying Chin, Yusairie Mohd, Nur Asyiqin Azman, and Zainiharyati Mohd Zain, Clinical Performance of Reverse Transcription Loop Mediated Isothermal Amplification COVID-19 Assay on Gold- nanoparticle-modified Screen-printed Carbon Electrode Using Differential Pulse Voltammetry, Sens. Mater., Vol. 35, No. 10, 2023, p. 4731-4750. |