Viewer specific sars cov 2

We performed nucleic acid extraction in a MicrolabStarlet IVD platform using the STARMag 96 × 4 universal Cartridge Kit (Seegene Inc., Seoul, South Korea). To detect SARS-CoV-2, we applied the Allplex™ SARS-CoV-2 Assay (Seegene Inc., Seoul, South Korea), a multiplex one-step rRT-PCR able to simultaneously detect four viral targets including the structural protein envelope (E) gene, the RNA-dependent RNA polymerase (RdRP) gene, the spike (S) gene, the nucleocapsid (N) gene, and an exogenous RNA-based internal control (IC). We targeted a conserved region in the structural protein envelope E-gene for pan-Sarbecovirus detection, RNA-dependent RNA polymerase (RdRP), and nucleocapsid (N) genes specific for SARS-CoV-2. For rRT-PCR, we employed the CFX96™ system (Bio-Rad Laboratories, Hercules, CA, United States). We analysed the results using Seegene Viewer-specific SARS-CoV-2 software (Seegene Inc., Seoul, South Korea). To establish a linear regression curve and obtain the concentration in copies/ml (inversely related to the cycle threshold value), the EDX SARS-CoV-2 Standard (Exact Diagnostics, TX, United States) containing 200,000 copies/mL of synthetic RNA transcripts from five gene targets (E, N, ORF1ab, RdRP and S Genes of SARS-CoV-2) was used.

We performed nucleic acid extraction in a MicrolabStarlet IVD platform using the STARMag 96 × 4 Universal Cartridge Kit (Seegene Inc, Seoul, South Korea). To detect SARS-CoV-2, we applied the Allplex™ SARS-CoV-2 Assay (Seegene Inc, Seoul, South Korea), a multiplex one-step rRT-PCR able to simultaneously detect four viral targets, including the structural protein envelope (E) gene, the RNA-dependent RNA polymerase (RdRP) gene, the spike (S) gene, the nucleocapsid (N) gene, and an exogenous RNA-based internal control (IC). This rRT-PCR step was run on a CFX96™ system (Bio-Rad Laboratories, Hercules, CA, USA), and the analysis was performed using Seegene Viewer-specific SARS-CoV-2 software (Seegene Inc, Seoul, South Korea), resulting in separate cycle threshold (Ct) values for the E and N genes and one combined Ct value for the RdRp and S genes (RdRp/S) in the FAM, Cal Red 610 and Quasar 670 channels, respectively. The HEX channel is used for internal control. Regarding interpretation of the results, according to the manufacturer’s instructions, Cts values ≤ 40 are considered detected, and Cts value > 40 or not applicable (N/A) are considered not detected.