Two extraction-free reverse transcription loop-mediated isothermal amplification assays for detection of SARS-CoV-2

BMC Infectious Diseases

Yee Leng Lee¹  Meng Yee Lai²  Fatma Diyana Mohd Bukhari²  Nur Zulaikha Zulkefli²  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, Kuala Lumpur, Malaysia;Department of Parasitology, Faculty of Medicine, University of Malaya, 50603, Kuala Lumpur, Malaysia;Department of Pathology, Hospital Sungai Buloh, Ministry of Health, Kuala Lumpur, Malaysia;Institute for Clinical Research, National Institutes of Health, Ministry of Health, Kuala Lumpur, Malaysia;Virology Unit, Infectious Disease Research Centre, Institute for Medical Research, National Institutes of Health, Ministry of Health, Kuala Lumpur, Malaysia;

关键词: SARS-CoV-2;    Isothermal detection;    Coronaviruses;    Rapid diagnosis;    RT-LAMP;   

DOI  :  10.1186/s12879-021-06876-0

来源: Springer

【 摘 要 】

BackgroundCurrent assays for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rely on time consuming, costly and laboratory based methods for virus isolation, purification and removing inhibitors. To address this limitation, we propose a simple method for testing RNA from nasopharyngeal swab samples that bypasses the RNA purification step.MethodsIn the current project, we have described two extraction-free reverse transcription loop-mediated isothermal amplification (RT-LAMP) assays for the detection of SARS-CoV-2 by using E gene and RdRp gene as the targets.ResultsHere, results showed that reverse transcription loop-mediated isothermal amplification assays with 88.4% sensitive (95% CI: 74.9–96.1%) and 67.4% sensitive (95% CI: 51.5–80.9%) for E gene and RdRp gene, respectively.ConclusionWithout the need of RNA purification, our developed RT-LAMP assays for direct detection of SARS-CoV-2 from nasopharyngeal swab samples could be turned into alternatives to qRT-PCR for rapid screening.

【 授权许可】

CC BY