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. 2020 Aug:129:104474.
doi: 10.1016/j.jcv.2020.104474. Epub 2020 May 28.

Validation and verification of the Abbott RealTime SARS-CoV-2 assay analytical and clinical performance

Emily Degli-Angeli  1 , Joan Dragavon  1 , Meei-Li Huang  2 , Danijela Lucic  3 , Gavin Cloherty  4 , Keith R Jerome  2 , Alexander L Greninger  2 , Robert W Coombs  5
Affiliations

Validation and verification of the Abbott RealTime SARS-CoV-2 assay analytical and clinical performance

Emily Degli-Angeli et al. J Clin Virol. 2020 Aug.

Abstract

Background: High-throughput assays for the SARS-CoV-2 virus are critical to increasing test capacity and slowing the spread of COVID-19. Abbott Molecular developed and received emergency use authorization (EUA) to deploy the new RealTime SARS-CoV-2 assay, run on the automated m2000sp/rt system.

Objective: To evaluate analytical and clinical performance of the RealTime SARS-CoV-2 assay compared to the SARS-CoV-2 CDC-based laboratory developed test (LDT) in clinical use by the University of Washington Clinical Virology Laboratory (UW Virology).

Methods: RealTime SARS-CoV-2 assay limit of detection (LOD) was evaluated by testing two dilution panels of 60 replicates each. Cross-reactivity was evaluated by testing 24 clinical samples positive for various non‒SARS-CoV-2 respiratory viruses. Clinical performance was evaluated using 30 positive and 30 negative SARS-CoV-2 clinical samples previously tested using the UW Virology SARS-CoV-2 LDT.

Results: Exceeding the 100 copies/mL LOD reported in the RealTime SARS-CoV-2 assay EUA product insert, 19 of 20 replicates were detected at 50 copies/mL and 16 of 20 replicates were detected at 25 copies/mL. All clinical samples positive for 24 non‒SARS-CoV-2 respiratory viruses were SARS-CoV-2 negative on the RealTime SARS-CoV-2 assay. The assay had high sensitivity (93%) and specificity (100%) for detecting SARS-CoV-2 in clinical samples. Two positive samples that tested negative with the RealTime SARS-CoV-2 assay had cycle numbers of 35.94 or greater and required dilution prior to testing. One of these samples was also inconclusive on the SARS-CoV-2 LDT.

Conclusion: The RealTime SARS-CoV-2 assay is acceptable for clinical use. With the high-throughput, fully automated m2000 system, this assay will accelerate the pace of SARS-CoV-2 testing.

Keywords: COVID-19; Coronavirus; EUA; High-throughput assay; PCR; SARS-CoV-2.

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Conflict of interest statement

Declaration of Competing Interest Alex L Greninger declares personal fees from Abbott Molecular, outside the scope of the submitted work. Danijela Lucic is an employee of Abbott Molecular. Gavin Cloherty is an employee of Abbott Diagnostics. Abbott Molecular provided the RealTime SARS-CoV-2 assays for use in the study but was not involved in data collection. The other authors have no declarations of interest.

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