RNA purification-free detection of SARS-CoV-2 using reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Tropical Medicine and Health

Yee Leng Lee¹  Meng Yee Lai²  Yee Ling Lau²  Ilyiana Ismail³  Nur Izati Mustapa³  Tuan Suhaila Tuan Soh³  Afifah Haji Hassan³  Kalaiarasu M. Peariasamy⁴  Jeyanthi Suppiah⁵  Ravindran Thayan⁵ 

[1]Clinical Research Centre, Hospital Sungai Buloh, Ministry of Health, Sungai Buloh, Selangor, Malaysia

[2]Department of Parasitology, Faculty of Medicine, University Malaya, 50603, Kuala Lumpur, Malaysia

[3]Department of Pathology, Hospital Sungai Buloh, Ministry of Health, Sungai Buloh, Selangor, Malaysia

[4]Institute for Clinical Research, National Institutes of Health, Ministry of Health, Shah Alam, Selangor, Malaysia

[5]Virology Unit, Infectious Disease Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health, Shah Alam, Selangor, Malaysia

关键词: COVID-19;    Nasopharyngeal swab;    SARS-CoV-2;    RT-LAMP;   

DOI  :  10.1186/s41182-021-00396-y

来源: Springer

【 摘 要 】

BackgroundCurrent diagnosis of SARS-CoV-2 infection relies on RNA purification prior to amplification. Typical extraction methods limit the processing speed and turnaround time for SARS-CoV-2 diagnostic testing.MethodsHere, we applied reverse transcription loop-mediated isothermal amplification directly onto human clinical swabs samples to amplify the RNA from SARS-CoV-2 swab samples after processing with chelating resin.ResultsBy testing our method on 64 samples, we managed to develop an RT-LAMP assay with 95.9% sensitivity (95% CI 86 to 99.5%) and 100% specificity (95% CI 78.2–100%).ConclusionThe entire process including sample processing can be completed in approximately 50 min. This method has promising potential to be applied as a fast, simple and inexpensive diagnostic tool for the detection of SARS-CoV-2.

【 授权许可】

CC BY