One Assay to Test Them All: Comparing Multiplex Assays for Expansion of Respiratory Virus Surveillance

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), TaqPathTM COVID-19, FluA/B, RSV Combo kit (Thermo Fisher Scientific, TP), and PowerChekTM 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.

The Role and Value of Professional Rapid Testing of Acute Respiratory Infections (ARIs) in Europe: A Special Focus on the Czech Republic, Poland, and Romania

“…Detects SARS-CoV-2 and influenza A and B [54,56] • Can be used for burden of disease surveillance [35,36,55].…”

Differential Diagnosis in the Management of Acute Respiratory Infections through Point-of-Care Rapid Testing in a Post-Pandemic Scenario in Latin America: Special Focus on COVID-19, Influenza, and Respiratory Syncytial Virus

“…These tests include various combinations such as influenza A, influenza B, and SARS-CoV-2; influenza A, influenza B, RSV, and SARS-CoV-2; and 20 of the most common respiratory viruses and bacteria causing upper respiratory illness. Information provided through these tests may be used for diagnosis, clinical management, and epidemiological surveillance (including the burden of disease virus surveillance) [215]. The second type is the rapid multiplex antigen test.…”