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The implementation of a rapid sample preparation method for the detection of SARS-CoV-2 in a diagnostic laboratory in South Africa

The SARS-CoV-2 pandemic has resulted in shortages of both critical reagents for nucleic acid purification and highly trained staff as supply chains are strained by high demand, public health measures and frequent quarantining and isolation of staff. This created the need for alternate workflows with...

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Autores principales: Marais, Gert, Naidoo, Michelle, Hsiao, Nei-yuan, Valley-Omar, Ziyaad, Smuts, Heidi, Hardie, Diana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7575110/
https://www.ncbi.nlm.nih.gov/pubmed/33079951
http://dx.doi.org/10.1371/journal.pone.0241029
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author Marais, Gert
Naidoo, Michelle
Hsiao, Nei-yuan
Valley-Omar, Ziyaad
Smuts, Heidi
Hardie, Diana
author_facet Marais, Gert
Naidoo, Michelle
Hsiao, Nei-yuan
Valley-Omar, Ziyaad
Smuts, Heidi
Hardie, Diana
author_sort Marais, Gert
collection PubMed
description The SARS-CoV-2 pandemic has resulted in shortages of both critical reagents for nucleic acid purification and highly trained staff as supply chains are strained by high demand, public health measures and frequent quarantining and isolation of staff. This created the need for alternate workflows with limited reliance on specialised reagents, equipment and staff. We present here the validation and implementation of such a workflow for preparing samples for downstream SARS-CoV-2 RT-PCR using liquid handling robots. The rapid sample preparation technique evaluated, which included sample centrifugation and heating prior to RT-PCR, showed a 97.37% (95% CI: 92.55–99.28%) positive percent agreement and 97.30% (95% CI: 90.67–99.52%) negative percent agreement compared to nucleic acid purification-based testing. This method was subsequently adopted as the primary sample preparation method in the Groote Schuur Hospital Virology Diagnostic Laboratory in Cape Town, South Africa.
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spelling pubmed-75751102020-10-26 The implementation of a rapid sample preparation method for the detection of SARS-CoV-2 in a diagnostic laboratory in South Africa Marais, Gert Naidoo, Michelle Hsiao, Nei-yuan Valley-Omar, Ziyaad Smuts, Heidi Hardie, Diana PLoS One Research Article The SARS-CoV-2 pandemic has resulted in shortages of both critical reagents for nucleic acid purification and highly trained staff as supply chains are strained by high demand, public health measures and frequent quarantining and isolation of staff. This created the need for alternate workflows with limited reliance on specialised reagents, equipment and staff. We present here the validation and implementation of such a workflow for preparing samples for downstream SARS-CoV-2 RT-PCR using liquid handling robots. The rapid sample preparation technique evaluated, which included sample centrifugation and heating prior to RT-PCR, showed a 97.37% (95% CI: 92.55–99.28%) positive percent agreement and 97.30% (95% CI: 90.67–99.52%) negative percent agreement compared to nucleic acid purification-based testing. This method was subsequently adopted as the primary sample preparation method in the Groote Schuur Hospital Virology Diagnostic Laboratory in Cape Town, South Africa. Public Library of Science 2020-10-20 /pmc/articles/PMC7575110/ /pubmed/33079951 http://dx.doi.org/10.1371/journal.pone.0241029 Text en © 2020 Marais et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Marais, Gert
Naidoo, Michelle
Hsiao, Nei-yuan
Valley-Omar, Ziyaad
Smuts, Heidi
Hardie, Diana
The implementation of a rapid sample preparation method for the detection of SARS-CoV-2 in a diagnostic laboratory in South Africa
title The implementation of a rapid sample preparation method for the detection of SARS-CoV-2 in a diagnostic laboratory in South Africa
title_full The implementation of a rapid sample preparation method for the detection of SARS-CoV-2 in a diagnostic laboratory in South Africa
title_fullStr The implementation of a rapid sample preparation method for the detection of SARS-CoV-2 in a diagnostic laboratory in South Africa
title_full_unstemmed The implementation of a rapid sample preparation method for the detection of SARS-CoV-2 in a diagnostic laboratory in South Africa
title_short The implementation of a rapid sample preparation method for the detection of SARS-CoV-2 in a diagnostic laboratory in South Africa
title_sort implementation of a rapid sample preparation method for the detection of sars-cov-2 in a diagnostic laboratory in south africa
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7575110/
https://www.ncbi.nlm.nih.gov/pubmed/33079951
http://dx.doi.org/10.1371/journal.pone.0241029
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