Amplirun total sars cov 2 control

Primers were purchased from SIGMA. Amplirun total SARS-CoV-2 Control and Amplirun MERS Coronavirus RNA Control were purchased from Vircell (Granada, Spain). Lucigen QuickExtract DNA(USA) kit was bought from Biosearch Technologies (Teddington, UK). COVID-19 negative saliva was purchased from Lee Biosolutions, Inc. (Maryland Heights, MO, USA).

Limit of detection (LOD) of the SORP triplex assay was determined using AmpliRun™ Total SARS-CoV-2 control (Vircell, Granada, Spain), an inactivated (swab) SARS-CoV-2 viral control preparation (4.3 × 10⁴ copies/mL). Post-reconstitution in sterile water as per the manufacturer's instructions, the SARS-CoV-2 inactivated virus control was extracted on the QIAsymphony (Qiagen) automated extraction platform, using the DSP Virus/Pathogen kit (Qiagen). Probit regression analysis to determine the 95% detection limit was carried out using the MedCalc Software package (MedCalc Software Ltd, Ostend, Belgium).
The analytical specificity of the SORP assay was evaluated using two methods: in silico analysis using BLAST analysis with common non-target organisms in the NCBI GenBank database (Table. 1_Suppl) and RT-qPCR analysis on a panel of non-target microorganisms. These non-target microorganisms were either positive patient specimens (nasopharyngeal/oropharyngeal swabs, bronchoalveolar lavage fluid, urine, saliva, and nasal washes), type strain cultures or commercial standards (Table. 2_Suppl). Nucleic acid was extracted from each individual organism using the QIAsymphony (Qiagen, Hilden, Germany) automated extraction platform.