Abstract
BackgroundHighly sensitive real-time reverse transcription polymerase chain reaction (RT-qPCR) methods have been developed for the detection of SARS-CoV-2. However, they are costly. Loop-mediated isothermal amplification (LAMP) assay has emerged as a novel alternative isothermal amplification method for the detection of nucleic acid.MethodsA rapid, sensitive and specific real-time reverse transcription LAMP (RT-LAMP) assay was developed for SARS-CoV-2 detection.ResultsThis assay detected one copy/reaction of SARS-CoV-2 RNA in 30 min. Both the clinical sensitivity and specificity of this assay were 100%. The RT-LAMP showed comparable performance with RT-qPCR. Combining simplicity and cost-effectiveness, this assay is therefore recommended for use in resource resource-limited settings.
Highlights
A novel coronavirus, SARS-CoV-2, was recently identified causing pneumonia in humans, termed coronavirus disease 2019 (COVID-19)
The analysis showed that the RT-Loop-mediated isothermal amplification (LAMP) developed is 100% specific and sensitive for the detection of SARS-CoV-2 with no false positives detected
The time required for confirmation of results by the reverse transcription LAMP (RT-LAMP) assay was less than 30 min, 2.5-fold faster than the time required by real-time PCR
Summary
A novel coronavirus, SARS-CoV-2, was recently identified causing pneumonia in humans, termed coronavirus disease 2019 (COVID-19). Several real-time reverse transcription polymerase chain reaction (RT-qPCR) methods have been developed and recommended by Centers for Disease Control of the United States (Centers for Disease Control, 2020) and WHO (World Health Organization, 2020b; Corman et al, 2020) for detection of SARS-like coronaviruses and specific detection of SARS-CoV-2. These methods are highly sensitive and specific but. In order to further reduce costs and enable detection by the naked eye, we used hydroxynaphthol blue (HNB) dye for the colorimetric detection of the amplification reaction To our knowledge, this is the first report of the detection of SARS-CoV-2 by real-time reverse transcription LAMP (RT-LAMP) assay with HNB. Combining simplicity and cost-effectiveness, this assay is recommended for use in resource resource-limited settings
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