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SARS-CoV-2 RNA extraction using magnetic beads for rapid large-scale testing by RT-qPCR and RT-LAMP
Steffen Klein; Thorsten G. Mueller; Dina Khalid; Vera Sonntag-Buck; Anke-Mareil Heuser; Baerbel Glass; Matthias Meurer; Ivonne Morales; Angelika Schillak; Andrew Freistaedter; Ina Ambiel; Sophie L. Winter; Liv Zimmermann; Tamara Naumoska; Felix Bubeck; Daniel Kirrmaier; Stephanie Ullrich; Isabel Barreto-Miranda; Simon Anders; Dirk Grimm; Paul Schnitzler; Michael Knop; Hans-Georg Kraeusslich; Viet Loan Dao Thi; Kathleen Boerner; Petr Chlanda.
Afiliación
  • Steffen Klein; Heidelberg University Hospital
  • Thorsten G. Mueller; Heidelberg University Hospital
  • Dina Khalid; Heidelberg University Hospital
  • Vera Sonntag-Buck; Heidelberg University Hospital
  • Anke-Mareil Heuser; Heidelberg University Hospital
  • Baerbel Glass; Heidelberg University Hospital
  • Matthias Meurer; Heidelberg University
  • Ivonne Morales; Heidelberg University Hospital
  • Angelika Schillak; Heidelberg University Hospital
  • Andrew Freistaedter; Heidelberg University Hospital
  • Ina Ambiel; Heidelberg University Hospital
  • Sophie L. Winter; Heidelberg University Hospital
  • Liv Zimmermann; Heidelberg University Hospital
  • Tamara Naumoska; Heidelberg University Hospital
  • Felix Bubeck; Heidelberg University Hospital
  • Daniel Kirrmaier; Heidelberg University Hospital
  • Stephanie Ullrich; Heidelberg University Hospital
  • Isabel Barreto-Miranda; Heidelberg University Hospital
  • Simon Anders; University of Heidelberg
  • Dirk Grimm; Heidelberg University Hospital
  • Paul Schnitzler; Heidelberg University Hospital
  • Michael Knop; Heidelberg University
  • Hans-Georg Kraeusslich; Heidelberg University Hospital
  • Viet Loan Dao Thi; Heidelberg University Hospital
  • Kathleen Boerner; Heidelberg University Hospital
  • Petr Chlanda; Heidelberg University Hospital
Preprint en En | PREPRINT-MEDRXIV | ID: ppmedrxiv-20147561
Artículo de revista
Un artículo publicado en revista científica está disponible y probablemente es basado en este preprint, por medio del reconocimiento de similitud realizado por una máquina. La confirmación humana aún está pendiente.
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ABSTRACT
Rapid large-scale testing is essential for controlling the ongoing pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The standard diagnostic pipeline for testing SARS-CoV-2 presence in patients with an ongoing infection is predominantly based on pharyngeal swabs, from which the viral RNA is extracted using commercial kits followed by reverse transcription and quantitative PCR detection. As a result of the large demand for testing, commercial RNA extraction kits may be limited and alternative, non-commercial protocols are needed. Here, we provide a magnetic bead RNA extraction protocol that is predominantly based on in-house made reagents and is performed in 96-well plates supporting large-scale testing. Magnetic bead RNA extraction was benchmarked against the commercial QIAcube extraction platform. Comparable viral RNA detection sensitivity and specificity were obtained by fluorescent and colorimetric RT-LAMP using N primers, as well as RT-qPCR using E gene primers showing that the here presented RNA extraction protocol can be combined with a variety of detection methods at high throughput. Importantly, the presented diagnostic workflow can be quickly set up in a laboratory without access to an automated pipetting robot.
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Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-MEDRXIV Tipo de estudio: Diagnostic_studies Idioma: En Año: 2020 Tipo del documento: Preprint
Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-MEDRXIV Tipo de estudio: Diagnostic_studies Idioma: En Año: 2020 Tipo del documento: Preprint