PLoS ONE · 2021 · 24 citations · 10 references
MedicineInfectious DiseasesMolecular BiologyDiagnostic MicrobiologyMolecular DiagnosticsVirologyInfectious Disease Molecular TestingMagnetic SeparationNucleic Acid ExtractionNucleic Acid PurificationRna-based DiagnosticsViral DiagnosticsAutomated Nucleic Acid ExtractionMagnetic Bead-based ExtractionRibonucleic Acid ExtractionRna ExtractionRobotic Liquid HandlingSars-cov-2 InfectionTissue Nucleic Acid ExtractionTotal Nucleic Acid Extraction
BACKGROUND: Two automatable in-house protocols for high-troughput RNA extraction from nasopharyngeal swabs for SARS-CoV-2 detection have been evaluated. METHODS: One hundred forty one SARS-CoV-2 positive samples were collected during a period of 10-days. In-house protocols were based on extraction with magnetic beads and designed to be used with either the Opentrons OT-2 (OT-2in-house) liquid handling robot or the MagMAXTM Express-96 system (MMin-house). Both protocols were tested in parallel with a commercial kit that uses the MagMAXTM system (MMkit). Nucleic acid extraction efficiencies were calculated from a SARS-CoV-2 DNA positive control. RESULTS: No significant differences were found between both in-house protocols and the commercial kit in their performance to detect positive samples. The MMkit was the most efficient although the MMin-house presented, in average, lower Cts than the other two. In-house protocols allowed to save between 350€ and 400€ for every 96 extracted samples compared to the commercial kit. CONCLUSION: The protocols described harness the use of easily available reagents and an open-source liquid handling system and are suitable for SARS-CoV-2 detection in high throughput facilities.
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SARS-CoV-2 detection by direct rRT-PCR without RNA extraction
Natacha Mérindol, Geneviève Pépin, Caroline Marchand et al. · Journal of Clinical Virology · 2020 · 112 citations · Full text