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Real-time reverse transcription loop-mediated isothermal amplification for rapid detection of SARS-CoV-2

Yee Ling Lau, Ilyiana Ismail, Nur Izati Mustapa, Meng Yee Lai, Tuan Suhaila Tuan Soh, Afifah Haji Hassan, Kalaiarasu M. Peariasamy, Yee Leng Lee, Yoong Min Chong, I‐Ching Sam, Pik Pin Goh

2020PeerJ47 citationsDOIOpen Access PDF

Abstract

BACKGROUND: Highly 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. METHODS: A rapid, sensitive and specific real-time reverse transcription LAMP (RT-LAMP) assay was developed for SARS-CoV-2 detection. RESULTS: This 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.

Topics & Concepts

Loop-mediated isothermal amplificationReverse Transcription Loop-mediated Isothermal AmplificationReverse transcriptaseReverse transcription polymerase chain reactionMolecular biologyNucleic acidReal-time polymerase chain reactionRecombinase Polymerase AmplificationSerial dilutionSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2)ChemistryPolymerase chain reactionBiologyVirologyCoronavirus disease 2019 (COVID-19)MedicineDNAGeneMessenger RNABiochemistryDiseasePathologyAlternative medicineInfectious disease (medical specialty)Biosensors and Analytical DetectionSARS-CoV-2 detection and testingAdvanced biosensing and bioanalysis techniques
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