A newly identified coronavirus, designated as severe acute respiratory syndrome coronavirus 2 (SARS CoV-2), has spread rapidly from its epicenter in China to more than 150 countries across six continents. In this study, we have designed three reverse-transcription loop-mediated isothermal amplification (RT-LAMP) primer sets to detect the RNA-dependent RNA polymerase (RdRP), Envelope (E) and Nucleocapsid protein (N) genes of SARS CoV-2. For one tube reaction, the detection limits for five combination SARS CoV-2 LAMP primer sets (RdRP/E, RdRP/N, E/N, RdRP/E/N and RdRP/N/Internal control (actin beta)) were evaluated with a clinical nasopharyngeal swab sample. Among the five combination, the RdRP/E and RdRP/N/IC multiplex LAMP assays showed low detection limits. The sensitivity and specificity of the RT-LAMP assay were evaluated and compared to that of the widely used Allplex™ 2019-nCoV Assay (Seegene, Inc., Seoul, South Korea) and PowerChek™ 2019-nCoV Real-time PCR kit (Kogenebiotech, Seoul, South Korea) for 130 clinical samples from 91 SARS CoV-2 patients and 162 NP specimens from individuals with (72) and without (90) viral respiratory infections. The multiplex RdRP (FAM)/N (CY5)/IC (Hex) RT-LAMP assay showed comparable sensitivities (RdRP: 93.85%, N: 94.62% and RdRP/N: 96.92%) to that of the Allplex™ 2019-nCoV Assay (100%) and superior to those of PowerChek™ 2019-nCoV Real-time PCR kit (RdRP: 92.31%, E: 93.85% and RdRP/E: 95.38%).
【초록키워드】 SARS CoV-2, respiratory infections, coronavirus, 2019-nCoV, severe acute respiratory syndrome Coronavirus, nucleocapsid protein, Spread, China, Nasopharyngeal swab, Protein, clinical samples, RT-LAMP, sensitivity, Viral, PCR, Sensitivity and specificity, Internal control, Loop-mediated isothermal amplification, LAMP, Patient, RdRP, RNA-dependent RNA polymerase, envelope, isothermal amplification, respiratory, multiplex, South Korea, Combination, Detection limit, (RdRp), acute respiratory syndrome, acute respiratory syndrome coronavirus, acute respiratory syndrome coronavirus 2, SARS CoV, individual, specimen, primer sets, limits, viral respiratory infections, nasopharyngeal swab sample, Actin, country, FIVE, clinical sample, detect, primer set, evaluated, comparable, Inc., 【제목키워드】 Diagnosis, multiplex, development,