Development of a reverse transcription loop‐mediated isothermal amplification assay with novel quantitative p H biosensor readout method for SARS ‐ C o V ‐2 detection

• Published in: APMIS : acta pathologica, microbiologica et immunologica Scandinavica microbiologica et immunologica Scandinavica, 2024-04
• Main Authors: Astari, Dian Ekayanti, Massi, Muhammad Nasrum, Masadah, Rina, Hardjo, Marhaen, Natzir, Rosdiana, Erlichster, Michael, Chana, Gursharan, Skafidas, Efstratios, Seraj, Zeba Islam, Elias, Sabrina M., Soraya, Gita Vita
• Format: Article
• Language: English
• ISSN: 0903-4641; 1600-0463
• DOI: 10.1111/apm.13415

Reverse transcription loop‐mediated isothermal amplification (RT‐LAMP) is a molecular amplification method that can detect SARS‐CoV‐2 in a shorter time than the current gold‐standard molecular diagnostic reverse transcription‐polymerase chain reaction (RT‐PCR). However, previously developed RT‐LAMP assays have mostly relied on highly subjective visual colorimetric interpretation. In this study, an RT‐LAMP assay was developed with quantitative measurement of reaction pH using a novel portable pH biosensor compared to qualitative colorimetric interpretation and gel electrophoresis, with 57 clinical COVID‐19 samples used for validation of the test. The LoD of the assay is 10 3 copies/μL. The highest sensitivity was found in the qualitative methods (93.75%), while the highest specificity and likelihood ratio was found in the pH sensor (87.5% and 6.72). On the sensor measurement, a significant difference (p