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. 2020 Aug 18;3(3):59.
doi: 10.3390/mps3030059.

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

Damien Coupeau  1 , Nicolas Burton  1 , Noémie Lejeune  1 , Suzanne Loret  1 , Astrid Petit  1 , Srdan Pejakovic  1 , Florian Poulain  1 , Laura Bonil  1 , Gabrielle Trozzi  1 , Laetitia Wiggers  1 , Kévin Willemart  1 , Emmanuel André  2   3 , Lies Laenen  2   3 , Lize Cuypers  2   3 , Marc Van Ranst  2 , Pierre Bogaerts  4 , Benoît Muylkens  1 , Nicolas Albert Gillet  1
Affiliations

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

Damien Coupeau et al. Methods Protoc. .

Abstract

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.

Keywords: COVID-19; RTqPCR; SARS-CoV-2; diagnosis.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1
Type of clinical specimen tubes.
Figure 2
Figure 2
RNA extraction workflow.
Figure 3
Figure 3
PCR loading form.
Figure 4
Figure 4
SARS-CoV-2 PCR amplification curves from successive 10-fold dilutions of a positive sample.
Figure 5
Figure 5
Validation process of the RT-qPCR data.
Figure 6
Figure 6
Timeline of the process.
Figure 7
Figure 7
Anticipated results.
See this image and copyright information in PMC

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