SARS-CoV-2 Detection for Diagnosis Purposes in the Setting of a Molecular Biology Research Lab.

COVID-19 RTqPCR SARS-CoV-2 diagnosis

Journal

Methods and protocols
ISSN: 2409-9279
Titre abrégé: Methods Protoc
Pays: Switzerland
ID NLM: 101720073

Informations de publication

Date de publication:
18 Aug 2020
Historique:
received: 02 07 2020
revised: 12 08 2020
accepted: 13 08 2020
entrez: 23 8 2020
pubmed: 23 8 2020
medline: 23 8 2020
Statut: epublish

Résumé

The emergence of the SARS-CoV-2 virus and the exponential growth of COVID-19 cases have created a major crisis for public health systems. The critical identification of contagious asymptomatic carriers requires the isolation of viral nucleic acids, reverse transcription, and amplification by PCR. However, the shortage of specific proprietary reagents or the lack of automated platforms have seriously hampered diagnostic throughput in many countries. Here, we provide a procedure for SARS-CoV-2 detection for diagnostic purposes from clinical samples in the setting of a basic research molecular biology lab. The procedure details the necessary steps for daily analysis of up to 500 clinical samples with a team composed of 12 experienced researchers. The protocol has been designed to rely on widely available reagents and devices, to cope with heterogeneous clinical specimens, to guarantee nucleic acid extraction from very scarce biological material, and to minimize the rate of false-negative results.

Identifiants

pubmed: 32824827
pii: mps3030059
doi: 10.3390/mps3030059
pmc: PMC7564796
pii:
doi:

Types de publication

Journal Article

Langues

eng

Subventions

Organisme : Fonds De La Recherche Scientifique - FNRS
ID : 34972507
Organisme : Fonds De La Recherche Scientifique - FNRS
ID : 31454280

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Auteurs

Damien Coupeau (D)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Nicolas Burton (N)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Noémie Lejeune (N)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Suzanne Loret (S)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Astrid Petit (A)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Srdan Pejakovic (S)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Florian Poulain (F)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Laura Bonil (L)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Gabrielle Trozzi (G)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Laetitia Wiggers (L)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Kévin Willemart (K)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Emmanuel André (E)

KU Leuven, Laboratory of Clinical Bacteriology and Mycology, campus Gasthuisberg, Herestraat 49, 3000 Leuven, Belgium.
Department of Laboratory Medicine, National Reference Center for Respiratory Pathogens, University Hospitals Leuven, Herestraat 49, 3000 Leuven, Belgium.

Lies Laenen (L)

KU Leuven, Laboratory of Clinical Bacteriology and Mycology, campus Gasthuisberg, Herestraat 49, 3000 Leuven, Belgium.
Department of Laboratory Medicine, National Reference Center for Respiratory Pathogens, University Hospitals Leuven, Herestraat 49, 3000 Leuven, Belgium.

Lize Cuypers (L)

KU Leuven, Laboratory of Clinical Bacteriology and Mycology, campus Gasthuisberg, Herestraat 49, 3000 Leuven, Belgium.
Department of Laboratory Medicine, National Reference Center for Respiratory Pathogens, University Hospitals Leuven, Herestraat 49, 3000 Leuven, Belgium.

Marc Van Ranst (M)

KU Leuven, Laboratory of Clinical Bacteriology and Mycology, campus Gasthuisberg, Herestraat 49, 3000 Leuven, Belgium.

Pierre Bogaerts (P)

CHU UCL NAMUR, Laboratory of microbiology and molecular biology, Site Godinne, 5530 Yvoir, Belgium.

Benoît Muylkens (B)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Nicolas Albert Gillet (NA)

Namur Research Institute for Life Sciences (NARILIS), Integrated Veterinary Research Unit (URVI), University of Namur, 61 rue de Bruxelles, 5000 Namur, Belgium.

Classifications MeSH