Colorimetric detection of SARS-CoV-2 by uracil-DNA glycosylase (UDG) reverse transcription loop-mediated isothermal amplification (RT-LAMP)
Meng Yee Lai, Fatma Diyana Mohd Bukhari, Nur Zulaikha Zulkefli, Ilyiana Ismail, Nur Izati Mustapa, Tuan Suhaila Tuan Soh, Afifah Haji Hassan, Kalaiarasu M. Peariasamy, Yee Leng Lee, Jeyanthi Suppiah, Ravindran Thayan, Yee Ling Lau
Abstract
OBJECTIVES: Preventing reverse transcription loop-mediated isothermal amplification (RT-LAMP) carryover contamination could be solved by adding deoxyuridine triphosphate (dUTP) and uracil-DNA glycosylase (UDG) into the reaction master mix. METHODS: RNA was extracted from nasopharyngeal swab samples by a simple RNA extraction method. RESULTS: Testing of 77 samples demonstrated 91.2% sensitivity (95% confidence interval [CI]: 78-98.2%) and 100% specificity (95% confidence interval: 92-100%) using UDG RT-LAMP. CONCLUSION: This colorimetric UDG RT-LAMP is a simple-to-use, fast, and easy-to-interpret method, which could serve as an alternative for diagnosis of SARS-CoV-2 infection, especially in remote hospitals and laboratories with under-equipped medical facilities.