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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Infectious Bacterial Diseases Research » Research » Publications at this Location » Publication #397141
Research Project: Development of Improved Diagnostic and Control Strategies for Brucellosis in Livestock and Wildlife

Location: Infectious Bacterial Diseases Research

Title: Evaluation of the PrimeFlow RNA assay as a method of detection of SARS-CoV-2 single and dual Infections

Author
item FALKENBERG, SHOLLIE - Auburn University
item Buckley, Alexandra
item Boggiatto, Paola

Submitted to: Proceedings of the National Academy of Sciences (PNAS)
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 11/6/2023
Publication Date: 11/24/2023
Citation: Falkenberg, S.M., Devries, A.C., Boggiatto, P.M. 2023. Evaluation of the PrimeFlow RNA assay as a method of detection of SARS-CoV-2 single and dual Infections. Proceedings of the National Academy of Sciences (PNAS). https://doi.org/10.1007/s10616-023-00608-9.
DOI: https://doi.org/10.1007/s10616-023-00608-9

Interpretive Summary: Emerging new variants of SARS-CoV-2 continue to show differences in virulence, host range, transmission rates, and mechanisms of immune escape. As these variants are found, animal models are used to understand these characteristics in order to better inform how the virus is changing. Development of a cell-based system in order to assess features of new variants would greatly speed up our ability to understand these changes, and how they may affect various populations. In this study, we used various cells lines from different animal species to assess three different SARS-CoV-2 variants, and determine whether this is a viable route to assess infection status. The results from this study will be of interest to scientific personnel working on SARS-CoV-2 in both human and animal model species.

Technical Abstract: Given the implications of increased transmissibility, virulence, host range, and immune escapes of emerging variants of SARS-CoV-2, developing models to better understand infection dynamics of variants is important, in particular, if certain variants may predominate in comparison to other variants in different populations. In this study, a PrimeFlow RNA assay using in-situ detection of SARS-CoV-2 single and dual infection in multiple cell lines was evaluated. The model included bovine, porcine, ovine, and cervid cell lines and a Vero cell line with different infection statuses including single infection with an Alpha, Delta, or Omicron variant and dual infection with Alpha/Omicron or Delta/Omicron. RNA probes specific for the Spike protein coding region, were designed (omicron or non-omicron specific). SARS-CoV-2 RNA was detected in the Vero E6 and fetal deer testicle cell lines by flow cytometry for all variants with increased detection in Vero E6 cells. Additionally, both Alpha and Delta predominated over Omicron in dual infections. This is the first report using the PrimeFlow assay for the detection of SARS-CoV-2 at the single-cell level as a model for competition of variants utilizing acute infection dynamics in cell culture.