Colorimetric detection of SARS-CoV-2 by uracil-DNA glycosylase (UDG) reverse transcription loop-mediated isothermal amplification (RT-LAMP)

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...

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Detalles Bibliográficos
Autores principales: Lai, Meng Yee, Bukhari, Fatma Diyana Mohd, Zulkefli, Nur Zulaikha, Ismail, Ilyiana, Mustapa, Nur Izati, Soh, Tuan Suhaila Tuan, Hassan, Afifah Haji, Peariasamy, Kalaiarasu M, Lee, Yee Leng, Suppiah, Jeyanthi, Thayan, Ravindran, Lau, Yee Ling
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Published by Elsevier Ltd on behalf of International Society for Infectious Diseases. 2022
Materias:
Short Communication
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9035611/
https://www.ncbi.nlm.nih.gov/pubmed/35472524
http://dx.doi.org/10.1016/j.ijid.2022.04.036
Descripción
Sumario: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.

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