4.7 Article

Fast SARS-CoV-2 detection by RT-qPCR in preheated nasopharyngeal swab samples

Journal

INTERNATIONAL JOURNAL OF INFECTIOUS DISEASES
Volume 97, Issue -, Pages 66-68

Publisher

ELSEVIER SCI LTD
DOI: 10.1016/j.ijid.2020.05.099

Keywords

COVID-19; SARS-CoV-2; diagnosis; sample treatment; RNA extraction; fast protocols

Funding

  1. Cabildo Insular de Tenerife [CGIEU0000219140]
  2. Instituto Tecnologico y de Energias Renovables (ITER) [OA17/008]
  3. Ministerio de Innovacion y Ciencia [RTI2018-093747-B-100, RTC-2017-6471-1]
  4. European Regional Development Fund (ERDF)
  5. Lab P2+ facility [UNLL10-3E-783]
  6. ERDF
  7. Fundacion CajaCanarias
  8. Spanish HIV/AIDS Research Network [RIS-RETIC, RD16/0025/0011]
  9. Instituto de Salud Carlos III

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Objectives: The gold-standard COVID-19 diagnosis relies on detecting SARS-CoV-2 using RNA purification and one-step retrotranscription and quantitative PCR (RT-qPCR). Based on the urgent need for highthroughput screening, we tested the performance of three alternative, simple and affordable protocols to rapidly detect SARS-CoV-2, bypassing the long and tedious RNA extraction step and reducing the time to viral detection. Methods: We evaluated three methods based on direct nasopharyngeal swab viral transmission medium (VTM) heating before the RT-qPCR: a) direct without additives; b) in a formamide-EDTA (FAE) buffer, c) in a RNAsnap (TM) buffer. Results: Although with a delay in cycle threshold compared to the gold-standard, we found consistent results in nasopharyngeal swab samples that were subject to a direct 70 degrees C incubation for 10 min. Conclusions: Our findings provide valuable options to overcome any supply chain issue and help to increase the throughput of diagnostic tests, thereby complementing standard diagnosis. (C) 2020 The Author(s). Published by Elsevier Ltd on behalf of International Society for Infectious Diseases. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-ncnd/4.0/).

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