Safe and effective pool testing for SARS-CoV-2 detection.


Journal

Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology
ISSN: 1873-5967
Titre abrégé: J Clin Virol
Pays: Netherlands
ID NLM: 9815671

Informations de publication

Date de publication:
12 2021
Historique:
received: 31 03 2021
revised: 01 08 2021
accepted: 19 10 2021
pubmed: 15 11 2021
medline: 15 12 2021
entrez: 14 11 2021
Statut: ppublish

Résumé

The global spread of SARS-CoV-2 is a serious public health issue. Large-scale surveillance screenings are crucial but can exceed test capacities. We (A) optimized test conditions and (B) implemented pool testing of respiratory swabs into SARS-CoV-2 diagnostics. (A) We determined the optimal pooling strategy and pool size. In addition, we measured the impact of vortexing prior to sample processing, compared a pipette-pooling method (by combining transport medium of several specimens) and a swab-pooling method (by combining several swabs into a test tube filled with PBS) as well as determined the sensitivities of three PCR assays. (B) Finally, we applied high-throughput pool testing for diagnostics. (A) In a low prevalence setting, we defined a preferable pool size of ten in a two-stage hierarchical pool testing strategy. Vortexing of swabs (n = 33) increased cellular yield by a factor of 2.34. By comparing Ct-values of 16 pools generated with two different pooling strategies, pipette-pooling was more efficient compared to swab-pooling. Measuring dilution series of 20 SARS-CoV-2 positive samples in three PCR assays simultaneously revealed detection rates of 85% (assay I), 50% (assay II), and 95% (assay III) at a 1:100 dilution. (B) We systematically pooled 55,690 samples in a period of 44 weeks resulting in a reduction of 47,369 PCR reactions. For implementing pooling strategies into high-throughput diagnostics, we recommend utilizing a pipette-pooling method, performing sensitivity validation of the PCR assays used, and vortexing swabs prior to analyses. Pool testing for SARS-CoV-2 detection is feasible and effective in a low prevalence setting.

Identifiants

pubmed: 34775143
pii: S1386-6532(21)00285-7
doi: 10.1016/j.jcv.2021.105018
pmc: PMC8552800
pii:
doi:

Substances chimiques

RNA, Viral 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

105018

Informations de copyright

Copyright © 2021. Published by Elsevier B.V.

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Auteurs

Marie Wunsch (M)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany; University of Cologne, Center for Molecular Medicine Cologne, Robert-Koch-Straße 21, 50931 Cologne, Germany.

Dominik Aschemeier (D)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany.

Eva Heger (E)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany.

Denise Ehrentraut (D)

Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Joseph-Stelzmann-Straße 26, 50931 Cologne, Germany.

Jan Krüger (J)

Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Joseph-Stelzmann-Straße 26, 50931 Cologne, Germany.

Martin Hufbauer (M)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany.

Adnan S Syed (AS)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany.

Gibran Horemheb-Rubio (G)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany; Department of Infectious Diseases, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Mexico City, 14080, Mexico.

Felix Dewald (F)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany; University of Cologne, Center for Molecular Medicine Cologne, Robert-Koch-Straße 21, 50931 Cologne, Germany.

Irina Fish (I)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany.

Maike Schlotz (M)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany; University of Cologne, Center for Molecular Medicine Cologne, Robert-Koch-Straße 21, 50931 Cologne, Germany.

Henning Gruell (H)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany; University of Cologne, Center for Molecular Medicine Cologne, Robert-Koch-Straße 21, 50931 Cologne, Germany.

Max Augustin (M)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Department I of Internal Medicine, Center for Integrated Oncology Aachen Bonn Cologne Duesseldorf, Kerpener Str. 62, 50937 Cologne, Germany.

Clara Lehmann (C)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Department I of Internal Medicine, Center for Integrated Oncology Aachen Bonn Cologne Duesseldorf, Kerpener Str. 62, 50937 Cologne, Germany.

Rolf Kaiser (R)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany.

Elena Knops (E)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany.

Steffi Silling (S)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany.

Florian Klein (F)

University of Cologne, Faculty of Medicine and University Hospital Cologne, Institute of Virology, Fürst-Pückler-Straße 56, 50935 Cologne, Germany; University of Cologne, Center for Molecular Medicine Cologne, Robert-Koch-Straße 21, 50931 Cologne, Germany. Electronic address: florian.klein@uk-koeln.de.

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