Clinical evaluation of a fully automated, laboratory-developed multiplex RT-PCR assay integrating dual-target SARS-CoV-2 and influenza A/B detection on a high-throughput platform

Dominik Nörz; Armin Hoffmann; Martin Aepfelbacher; Susanne Pfefferle; Marc Lütgehetmann

doi:10.1099/jmm.0.001295

Volume 70, Issue 2

Short Communication

Open Access

Clinical evaluation of a fully automated, laboratory-developed multiplex RT-PCR assay integrating dual-target SARS-CoV-2 and influenza A/B detection on a high-throughput platform

Dominik Nörz¹, Armin Hoffmann¹, Martin Aepfelbacher¹, Susanne Pfefferle^1,† and Marc Lütgehetmann^1,†
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Affiliations: ¹ University Medical Center Hamburg-Eppendorf (UKE), Institute of Medical Microbiology, Virology and Hygiene, Hamburg, Germany
*Correspondence: Marc Lütgehetmann, [email protected]

† These authors contributed equally to this work
Published: 06 January 2021 https://doi.org/10.1099/jmm.0.001295

Abstract

Introduction. Laboratories worldwide are facing high demand for molecular testing during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, which might be further aggravated by the upcoming influenza season in the northern hemisphere.

Gap Statement. Given that the symptoms of influenza are largely indistinguishable from those of coronavirus disease 2019 (COVID-19), both SARS-CoV-2 and the influenza viruses require concurrent testing by RT-PCR in patients presenting with symptoms of respiratory tract infection.

Aim. We adapted and evaluated a laboratory-developed multiplex RT-PCR assay for simultaneous detection of SARS-CoV-2 (dual target), influenza A and influenza B (SC2/InflA/InflB-UCT) on a fully automated high-throughput system (cobas6800).

Methodology. Analytical performance was assessed by serial dilution of quantified reference material and cell culture stocks in transport medium, including pretreatment for chemical inactivation. For clinical evaluation, residual portions of 164 predetermined patient samples containing SARS-CoV-2 (n=52), influenza A (n=43) or influenza B (n=19), as well as a set of negative samples, were subjected to the novel multiplex assay.

Results. The assay demonstrated comparable analytical performance to currently available commercial tests, with limits of detection of 94.9 cp ml−1 for SARS-CoV-2, 14.6 cp ml−1 for influenza A and 422.3 cp ml−1 for influenza B. Clinical evaluation showed excellent agreement with the comparator assays (sensitivity of 98.1, 97.7 and 100 % for Sars-CoV-2 and influenza A and B, respectively).

Conclusion. The SC2/InflA/InflB-UCT allows for efficient high-throughput testing for all three pathogens and thus provides streamlined diagnostics while conserving resources during the influenza season.

Received: 07/09/2020
Accepted: 28/11/2020
Published Online: 06/01/2021

Keyword(s): cobas6800 , influenza , molecular diagnostics , multiplex , PCR and SARS-CoV-2

This is an open-access article distributed under the terms of the Creative Commons Attribution NonCommercial License. The Microbiology Society waived the open access fees for this article.

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Clinical evaluation of a fully automated, laboratory-developed multiplex RT-PCR assay integrating dual-target SARS-CoV-2 and influenza A/B detection on a high-throughput platform

J. Med. Microbiol. 70, 001295 (2021); https://doi.org/10.1099/jmm.0.001295

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Volume 70, Issue 2

Short Communication

Open Access

Clinical evaluation of a fully automated, laboratory-developed multiplex RT-PCR assay integrating dual-target SARS-CoV-2 and influenza A/B detection on a high-throughput platform

Abstract

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