Dry loop-mediated isothermal amplification assay for detection of SARS-CoV-2 from clinical specimens

Autor: Yuki Higashimoto, Masaru Ihira, Yoshiki Kawamura, Masato Inaba, Kazuya Shirato, Tadaki Suzuki, Hideki Hasegawa, Tsutomu Kageyama, Yohei Doi, Tadayoshi Hata, Tetsushi Yoshikawa
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: Fujita Medical Journal, Vol 9, Iss 2, Pp 84-89 (2023)
Druh dokumentu: article
ISSN: 2189-7247
2189-7255
DOI: 10.20407/fmj.2022-003
Popis: Objectives: To establish a point-of-care test for coronavirus disease 2019 (COVID-19), we developed a dry loop-mediated isothermal amplification (LAMP) method to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA. Methods: We carried out reverse transcription (RT)-LAMP using the Loopamp SARS-CoV-2 Detection kit (Eiken Chemical, Tokyo, Japan). The entire mixture, except for the primers, is dried and immobilized inside the tube lid. Results: To determine the specificity of the kit, 22 viruses associated with respiratory infections, including SARS-CoV-2, were tested. The sensitivity of this assay, determined by either a real-time turbidity assay or colorimetric change of the reaction mixture, as evaluated by the naked eye or under illumination with ultraviolet light, was 10 copies/reaction. No LAMP product was detected in reactions performed with RNA from any pathogens other than SARS-CoV-2. After completing an initial validation analysis, we analyzed 24 nasopharyngeal swab specimens collected from patients suspected to have COVID-19. Of the 24 samples, 19 (79.2%) were determined by real-time RT-PCR analysis as being positive for SARS-CoV-2 RNA. Using the Loopamp SARS-CoV-2 Detection kit, we detected SARS-CoV-2 RNA in 15 (62.5%) of the 24 samples. Thus, the sensitivity, specificity, positive predictive value, and negative predictive values of the Loopamp 2019-CoV-2 detection reagent kit were 78.9%, 100%, 100%, and 55.6%, respectively. Conclusions: The dry LAMP method for detecting SARS-CoV-2 RNA is fast and easy to use, and its reagents can be stored at 4°C, solving the cold chain problem; thus, it represents a promising tool for COVID-19 diagnosis in developing countries.
Databáze: Directory of Open Access Journals
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