Abstract
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.
Highlights
Current 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
These genes have been used in reverse transcription loop-mediated isothermal amplification (RT-loop-mediated isothermal amplification (LAMP)) as well [5, 7,8,9,10,11], most of these methods required RNA extraction and purification using commercial kits which is costly and time consuming
The RT-LAMP using E gene was 88.4% sensitive and 86.7% specific
Summary
Current 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. Laboratory screening test of SARS-CoV-2 using RT-PCR exploits a few different types of detection gene including envelope (E) gene, nucleocapside (N) gene [2, 8] and RNA dependent RNA Polymerase (RdRp) gene [2] These genes are significant for overall function of coronavirus and conserved [9]. These genes have been used in RT-LAMP as well [5, 7,8,9,10,11], most of these methods required RNA extraction and purification using commercial kits which is costly and time consuming
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