Increasing diagnostic possibilities using the geneLEAD VIII platform for detection of SARS-CoV-2

Stefan A Boers,Bas C Mourik,Mario J.A.W.M Van Bussel,Caroline S De Brouwer,Els Wessels,Eric C.J Claas

doi:10.1016/j.jviromet.2021.114291

Stefan A Boers, Bas C Mourik + Show 4 more

Open Access

https://doi.org/10.1016/j.jviromet.2021.114291

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Abstract

At the time SARS-CoV-2 was identified as the cause of coronavirus disease 2019 (COVID-19) no in vitro diagnostic (IVD) tests were available since it was a new virus. Very shortly after the release of the genomic sequence of SARS-CoV-2, laboratory-developed tests (LDTs) were developed, made available and implemented in several laboratories in the Netherlands and globally. In this study, the performance of an E-gene Sarbeco specific real-time reverse-transcriptase PCR (RT-PCR) was verified on the open modus of the geneLEAD VIII sample-to-answer platform. The results obtained from 134 clinical samples, of which 63 had been tested positive, showed almost complete concordance compared to the same PCR on the routine diagnostic systems and that was validated according to the national reference standard. The only discordant sample tested positive using the routine diagnostic workflow with a cycle threshold (CT) value of 37.7, while the sample tested negative using the geneLEAD VIII workflow. In addition, good performance was achieved in analyzing a blinded SARS-CoV-2 external quality assurance (EQA) panel. Implementation of the geneLEAD VIII platform as routine diagnostic tool resulted in testing 871 clinical samples with 115 positive results. In conclusion, the geneLEAD VIII SARS-CoV-2 workflow presented in this study showed excellent diagnostic performance and with a rapid turnaround time of approximately two hours it proved a valuable option for STAT SARS-CoV-2 testing in the absence of (rapid, CE-IVD) point-of-care testing platforms.

Highlights

Molecular diagnostics is the gold standard for severe acute respira tory syndrome coronavirus 2 (SARS-CoV-2) detection
The geneLEAD VIII workflow consisted of an on-board nucleic acid (NA) extraction followed by reverse-transcriptase PCR (RT-PCR)
* The viral load of three SARS-CoV-2 positive throat/nasopharyngeal swabs and three SARS-CoV-2 positive sputum samples was too high for an automatic cycle threshold (CT) calculation by the geneLEAD VIII software and required visual evaluation of the amplification signal for a final result

Summary

Introduction

Molecular diagnostics is the gold standard for severe acute respira tory syndrome coronavirus 2 (SARS-CoV-2) detection. The geneLEAD VIII workflow consisted of an on-board NA extraction followed by RT-PCR. * The viral load of three SARS-CoV-2 positive throat/nasopharyngeal swabs and three SARS-CoV-2 positive sputum samples was too high for an automatic CT calculation by the geneLEAD VIII software and required visual evaluation of the amplification signal for a final result.

Results

Conclusion