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[Preprint]. 2023 Jan 22:2023.01.19.23284806.
doi: 10.1101/2023.01.19.23284806.

ONE ASSAY TO TEST THEM ALL: COMPARING MULTIPLEX ASSAYS FOR EXPANSION OF RESPIRATORY VIRUS SURVEILLANCE

Narjis Boukli  1 , Claude Flamand  2 , Kim Lay Chea  1 , Leangyi Heng  1 , Seangmai Keo  1 , Kimhoung Sour  1 , Sophea In  1 , Panha Chhim  1 , Bunthea Chhor  1 , Lomor Kruy  1 , Jelena D M Feenstra  3 , Manoj Gandhi  3 , Obiageli Okafor  3 , Camilla Ulekliev  3 , Heidi Auerswald  1 , Viseth Srey Horm  1 , Erik A Karlsson  1
Affiliations

ONE ASSAY TO TEST THEM ALL: COMPARING MULTIPLEX ASSAYS FOR EXPANSION OF RESPIRATORY VIRUS SURVEILLANCE

Narjis Boukli et al. medRxiv. .

Update in

Abstract

Background: Molecular multiplex assays (MPAs) for simultaneous detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), influenza and respiratory syncytial virus (RSV) in a single RT-PCR reaction reduce time and increase efficiency to identify multiple pathogens with overlapping clinical presentation but different treatments or public health implications.

Methods: Clinical performance of XpertXpress ® SARS-CoV-2/Flu/RSV (Cepheid, GX), TaqPath™ COVID-19, FluA/B, RSV Combo kit (Thermo Fisher Scientific, TP), and PowerChek™ SARS-CoV-2/Influenza A&B/RSV Multiplex RT-PCR kit II (KogeneBiotech, PC) was compared to individual Standards of Care (SoC). Thirteen isolates of SARS-CoV-2, human seasonal influenza, and avian influenza served to assess limit of detection (LoD). Then, positive and negative residual nasopharyngeal specimens, collected under public health surveillance and pandemic response served for evaluation. Subsequently, comparison of effectiveness was assessed.

Results: The three MPAs confidently detect all lineages of SARS-CoV-2 and influenza viruses. MPA-LoDs vary from 1-2 Log10 differences from SoC depending on assay and strain. Clinical evaluation resulted in overall agreement between 97% and 100%, demonstrating a high accuracy to detect all targets. Existing differences in costs, testing burden and implementation constraints influence the choice in primary or community settings.

Conclusion: TP, PC and GX, reliably detect SARS-CoV-2, influenza and RSV simultaneously, with reduced time-to-results and simplified workflows. MPAs have the potential to enhancediagnostics, surveillance system, and epidemic response to drive policy on prevention and control of viral respiratory infections.

Importance: Viral respiratory infections represent a major burden globally, weighed down by the COVID-19 pandemic, and threatened by spillover of novel zoonotic influenza viruses. Since respiratory infections share clinical presentations, identification of the causing agent for patient care and public health measures requires laboratory testing for several pathogens, including potential zoonotic spillovers. Simultaneous detection of SARS-CoV-2, influenza, and RSV in a single RT-PCR accelerates time from sampling to diagnosis, preserve consumables, and streamline human resources to respond to other endemic or emerging pathogens. Multiplex assays have the potential to sustain and even expand surveillance systems, can utilize capacity/capability developed during the COVID-19 pandemic worldwide, thereby strengthening epidemic/pandemic preparedness, prevention, and response.

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Conflict of interest statement

CONFLICT OF INTEREST

This work was supported by Thermo Fisher Scientific, who loaned the QuantStudio 5™ RT-PCR, 96 well, 0.2 mL instrument (Applied Biosystems) and laptop to IPC for the purpose of the study, provided TaqPath™ COVID-19, FluA/B, RSV Combo Kits, and co-authors included were involved in the study design, analysis and interpretation of TaqPath results and reviewed the report.

Figures

FIGURE 1:
FIGURE 1:
Distribution of cycle threshold (Ct)-values (median; min-max) in clinical samples according to each RT-PCR assay. Standards of Care (SoC) are displayed in dark orange, Thermofisher TaqPath™ COVID-19, FluA/B, RSV Combo Kit (TP) in light blue, Kogene PowerChek™ SARS-CoV-2, Influenza A&B, RSV Multiplex Real-time PCR Kit II (PC) in light orange, Cephied Xpert® Xpress SARS-CoV-2, Flu, RSV Kit (GX) in grey. Ct values are displayed for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), respiratory
See this image and copyright information in PMC

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