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Fast Detection of SARS-CoV-2 RNA Directly from Respiratory Samples Using a Loop-Mediated Isothermal Amplification (LAMP) Test.
Anastasiou, Olympia E; Holtkamp, Caroline; Schäfer, Miriam; Schön, Frieda; Eis-Hübinger, Anna Maria; Krumbholz, Andi.
  • Anastasiou OE; Institute for Virology, University Hospital Essen, University of Duisburg-Essen, 45147 Essen, Germany.
  • Holtkamp C; Institute for Virology, University Hospital Essen, University of Duisburg-Essen, 45147 Essen, Germany.
  • Schäfer M; Labor Dr. Krause und Kollegen MVZ GmbH, 24106 Kiel, Germany.
  • Schön F; Labor Dr. Krause und Kollegen MVZ GmbH, 24106 Kiel, Germany.
  • Eis-Hübinger AM; Institute of Virology, Medical Faculty, University Hospital Bonn, 53127 Bonn, Germany.
  • Krumbholz A; Labor Dr. Krause und Kollegen MVZ GmbH, 24106 Kiel, Germany.
Viruses ; 13(5)2021 04 29.
Article in English | MEDLINE | ID: covidwho-1217117
ABSTRACT
The availability of simple SARS-CoV-2 detection methods is crucial to contain the COVID-19 pandemic. This study examined whether a commercial LAMP assay can reliably detect SARS-CoV-2 genomes directly in respiratory samples without having to extract nucleic acids (NA) beforehand. Nasopharyngeal swabs (NPS, n = 220) were tested by real-time reverse transcription (RT)-PCR and with the LAMP assay. For RT-PCR, NA were investigated. For LAMP, NA from 26 NPS in viral transport medium (VTM) were tested. The other 194 NPS were analyzed directly without prior NA extraction (140 samples in VTM; 54 dry swab samples stirred in phosphate buffered saline). Ten NPS were tested directly by LAMP using a sous-vide cooking unit. The isothermal assay demonstrated excellent specificity (100%) but moderate sensitivity (68.8%), with a positive predictive value of 1 and a negative predictive value of 0.65 for direct testing of NPS in VTM. The use of dry swabs, even without NA extraction, improved the analytical sensitivity; up to 6% of samples showed signs of inhibition. LAMP could be performed successfully with a sous-vide cooking unit. This technique is very fast, requires little laboratory resources, and can replace rapid antigen tests or verify reactive rapid tests on-site.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: DNA, Viral / Nasopharynx / Nucleic Acid Amplification Techniques / Molecular Diagnostic Techniques / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Year: 2021 Document Type: Article Affiliation country: V13050801

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Full text: Available Collection: International databases Database: MEDLINE Main subject: DNA, Viral / Nasopharynx / Nucleic Acid Amplification Techniques / Molecular Diagnostic Techniques / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Year: 2021 Document Type: Article Affiliation country: V13050801