Miracle autoxt automated nucleic acid extraction system

An experienced phlebotomy nurse collected blood. A total of 2 mL of blood was collected from the cubital vein by using a gold-top (serum separator) tube. Samples were stored in a sealed biohazard bag and transported at 4 °C in a cool transport container to the Khalifa University Center for Biotechnology’s laboratory for a second confirmatory testing for SARS-CoV-2. Viral RNA was extracted from swab by using the Miracle-AutoXT Automated Nucleic Acid Extraction System (iNtRON Biotechnology, Seoul, South Korea). Genesig from the Primerdesign reverse transcription-PCR COVID-19 detection kit (Watchmoor Point, UK) was used to quantify the viral RNA. PCRs were performed according to the manufacturer’s instructions. Quantitative reverse transcription-PCR was performed using the Magnetic Induction Cycler PCR Machine (MiC) (Bio Molecular Systems, Queensland, Australia).

Viral RNA was extracted using a Miracle-AutoXT Automated Nucleic Acid Extraction System (iNtRON Biotechnology, Seongnam, Korea). To identify subtypes, qRT-PCR was conducted using the TOPreal™ One-step RT qPCR Kit (Enzynomics, Daejeon, Korea) with influenza-specific primers as previously described [35 (link)]. Species identification was performed using mitochondrial cytochrome C oxidase I mitochondrial gene in the DNA of fecal samples, as previously described [36 (link)].

Viral RNA from nasal swabs was extracted using the Miracle-AutoXT Automated Nucleic Acid Extraction System (iNtRON Biotechnology Inc., South Korea), and viral RNA from fecal samples was extracted using DNeasy PowerLyzer PowerSoil Kit (Qiagen Ltd., GmbH, Germany) following the manufacturer’s instruction. Genes from the PrimerDesign RT-PCR COVID-19 detection kit (PrimerDesign Ltd.) were used for the quantification of the viral RNA in nasal swabs and fecal samples. The primers provided in the kit target the RdRp gene. Internal extraction control primers were also provided to detect the exogenous source of the RNA template added during the extraction step. The PCRs were performed according to the manufacturer protocol using the Magnetic Induction Cycler (MiC) PCR Machine (Bio Molecular Systems, QLD, Australia).