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Critical Aspects Concerning the Development of a Pooling Approach for SARS-CoV-2 Diagnosis Using Large-Scale PCR Testing

The primary approach to controlling the spread of the pandemic SARS-CoV-2 is to diagnose and isolate the infected people quickly. Our paper aimed to investigate the efficiency and the reliability of a hierarchical pooling approach for large-scale PCR testing for SARS-CoV-2 diagnosis. To identify the...

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Autores principales: Cruceriu, Daniel, Baldasici, Oana, Balacescu, Loredana, Gligor-Popa, Stefana, Flonta, Mirela, Man, Milena A., Visan, Simona, Vlad, Catalin, Trifa, Adrian P., Balacescu, Ovidiu, Achimas-Cadariu, Patriciu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8152296/
https://www.ncbi.nlm.nih.gov/pubmed/34067983
http://dx.doi.org/10.3390/v13050902
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author Cruceriu, Daniel
Baldasici, Oana
Balacescu, Loredana
Gligor-Popa, Stefana
Flonta, Mirela
Man, Milena A.
Visan, Simona
Vlad, Catalin
Trifa, Adrian P.
Balacescu, Ovidiu
Achimas-Cadariu, Patriciu
author_facet Cruceriu, Daniel
Baldasici, Oana
Balacescu, Loredana
Gligor-Popa, Stefana
Flonta, Mirela
Man, Milena A.
Visan, Simona
Vlad, Catalin
Trifa, Adrian P.
Balacescu, Ovidiu
Achimas-Cadariu, Patriciu
author_sort Cruceriu, Daniel
collection PubMed
description The primary approach to controlling the spread of the pandemic SARS-CoV-2 is to diagnose and isolate the infected people quickly. Our paper aimed to investigate the efficiency and the reliability of a hierarchical pooling approach for large-scale PCR testing for SARS-CoV-2 diagnosis. To identify the best conditions for the pooling approach for SARS-CoV-2 diagnosis by RT-qPCR, we investigated four manual methods for both RNA extraction and PCR assessment targeting one or more of the RdRp, N, S, and ORF1a genes, by using two PCR devices and an automated flux for SARS-CoV-2 detection. We determined the most efficient and accurate diagnostic assay, taking into account multiple parameters. The optimal pool size calculation included the prevalence of SARS-CoV-2, the assay sensitivity of 95%, an assay specificity of 100%, and a range of pool sizes of 5 to 15 samples. Our investigation revealed that the most efficient and accurate procedure for detecting the SARS-CoV-2 has a detection limit of 2.5 copies/PCR reaction. This pooling approach proved to be efficient and accurate in detecting SARS-CoV-2 for all samples with individual quantification cycle (Cq) values lower than 35, accounting for more than 94% of all positive specimens. Our data could serve as a comprehensive practical guide for SARS-CoV-2 diagnostic centers planning to address such a pooling strategy.
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spelling pubmed-81522962021-05-27 Critical Aspects Concerning the Development of a Pooling Approach for SARS-CoV-2 Diagnosis Using Large-Scale PCR Testing Cruceriu, Daniel Baldasici, Oana Balacescu, Loredana Gligor-Popa, Stefana Flonta, Mirela Man, Milena A. Visan, Simona Vlad, Catalin Trifa, Adrian P. Balacescu, Ovidiu Achimas-Cadariu, Patriciu Viruses Article The primary approach to controlling the spread of the pandemic SARS-CoV-2 is to diagnose and isolate the infected people quickly. Our paper aimed to investigate the efficiency and the reliability of a hierarchical pooling approach for large-scale PCR testing for SARS-CoV-2 diagnosis. To identify the best conditions for the pooling approach for SARS-CoV-2 diagnosis by RT-qPCR, we investigated four manual methods for both RNA extraction and PCR assessment targeting one or more of the RdRp, N, S, and ORF1a genes, by using two PCR devices and an automated flux for SARS-CoV-2 detection. We determined the most efficient and accurate diagnostic assay, taking into account multiple parameters. The optimal pool size calculation included the prevalence of SARS-CoV-2, the assay sensitivity of 95%, an assay specificity of 100%, and a range of pool sizes of 5 to 15 samples. Our investigation revealed that the most efficient and accurate procedure for detecting the SARS-CoV-2 has a detection limit of 2.5 copies/PCR reaction. This pooling approach proved to be efficient and accurate in detecting SARS-CoV-2 for all samples with individual quantification cycle (Cq) values lower than 35, accounting for more than 94% of all positive specimens. Our data could serve as a comprehensive practical guide for SARS-CoV-2 diagnostic centers planning to address such a pooling strategy. MDPI 2021-05-13 /pmc/articles/PMC8152296/ /pubmed/34067983 http://dx.doi.org/10.3390/v13050902 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Cruceriu, Daniel
Baldasici, Oana
Balacescu, Loredana
Gligor-Popa, Stefana
Flonta, Mirela
Man, Milena A.
Visan, Simona
Vlad, Catalin
Trifa, Adrian P.
Balacescu, Ovidiu
Achimas-Cadariu, Patriciu
Critical Aspects Concerning the Development of a Pooling Approach for SARS-CoV-2 Diagnosis Using Large-Scale PCR Testing
title Critical Aspects Concerning the Development of a Pooling Approach for SARS-CoV-2 Diagnosis Using Large-Scale PCR Testing
title_full Critical Aspects Concerning the Development of a Pooling Approach for SARS-CoV-2 Diagnosis Using Large-Scale PCR Testing
title_fullStr Critical Aspects Concerning the Development of a Pooling Approach for SARS-CoV-2 Diagnosis Using Large-Scale PCR Testing
title_full_unstemmed Critical Aspects Concerning the Development of a Pooling Approach for SARS-CoV-2 Diagnosis Using Large-Scale PCR Testing
title_short Critical Aspects Concerning the Development of a Pooling Approach for SARS-CoV-2 Diagnosis Using Large-Scale PCR Testing
title_sort critical aspects concerning the development of a pooling approach for sars-cov-2 diagnosis using large-scale pcr testing
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8152296/
https://www.ncbi.nlm.nih.gov/pubmed/34067983
http://dx.doi.org/10.3390/v13050902
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