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

Document Type

Article

Publication Date

1-4-2022

Abstract

Background: Current 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. Methods: Here, 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. Results: By 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). Conclusion: The 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. © 2022, The Author(s).

Keywords

Virus RNA, Article, Controlled study, Geobacillus stearothermophilus, Human, Isothermal amplification, Loop mediated isothermal amplification, Nasopharyngeal swab, Nonhuman, Oropharyngeal swab, Reverse transcription loop mediated isothermal amplification, RNA purification, Sensitivity and specificity, Severe acute respiratory syndrome coronavirus 2, Turnaround time, Virus nucleocapsid

Divisions

Parasit

Funders

Prototype Research Grant Scheme (PRGS) from the Ministry of Education, Malaysia [Grant No: PR001-2020B (PRGS/2/2020/SKK09/UM/02/1)]

Publication Title

Tropical Medicine and Health

Volume

50

Issue

1

Publisher

BioMed Central Ltd

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