Direct Detection of SARS-CoV-2 Using Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Without RNA Purification

Autor: Kalaiarasu M. Peariasamy, Ravindran Thayan, Ilyiana Ismail, Jeyanthi Suppiah, Yee Leng Lee, Afifah Haji Hassan, Fatma Diyana Mohd Bukhari, Meng Yee Lai, Nur Izati Mustapa, Yee Ling Lau, Tuan Suhaila Tuan Soh, Nur Zulaikha Zulkefli
Rok vydání: 2021
Předmět:
Zdroj: SSRN Electronic Journal.
ISSN: 1556-5068
2041-8418
Popis: Coronavirus disease 2019 (COVID-19) is an acute respiratory disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) which has spread globally. Shortage of nucleic acid extraction kits may delay the speed of diagnosis. In order to address this need, we propose a simple method for direct RNA extraction from nasopharyngeal swab samples without RNA purification. Here, we describe a reverse transcription loop-mediated isothermal amplification technique with 95.4% sensitivity (95% CI: 84.2-99.4%) and 100% specificity (95% CI: 78-100%). Combined with a simple RNA extraction step, this optimized RT-LAMP method is able to amplify SARS-CoV-2 directly from clinical nasopharyngeal swab samples in viral transport media in 50 minutes. This method provides an opportunity to facilitate SARS-CoV-2 detection especially in resource limited settings where nucleic acid extraction kits are few. Funding: This study was supported by Prototype Research Grant Scheme (PRGS), PR001-2020B (PRGS/2/2020/SKK09/UM/02/1) from the Ministry of Education, Malaysia. Declaration of Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this manuscript. Ethics Approval Statement: Ethical approvals for this study was obtained from UMMC 85 Medical Ethics Committee (202041-8418) and Medical Research Ethics Committee 86 (MREC) Ministry of Health Malaysia (NMRR-20-2344-56994).
Databáze: OpenAIRE
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