Development of an RNA Strand-Specific Hybridization Assay To Differentiate Replicating versus Nonreplicating Influenza A Viruses

Author:

Yang Genyan1,Hodges Erin N.1,Winter Jörn1,Zanders Natosha1,Shcherbik Svetlana1,Bousse Tatiana1,Murray Janna R.1,Muraduzzaman A. K. M.2,Rahman Mahbubur2,Alamgir A. S. M.2,Flora Meerjady Sabrina2,Blanton Lenee1,Barnes John R.1,Wentworth David E.1,Davis C. Todd1

Affiliation:

1. Influenza Division, National Center for Immunization and Respiratory Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA

2. Institute of Epidemiology, Disease Control and Research, Dhaka, Bangladesh

Abstract

Replication of influenza A virus (IAV) from negative-sense viral RNA (vRNA) requires the generation of positive-sense RNA (+RNA). Most molecular assays, such as conventional real-time reverse transcriptase PCR (rRT-PCR), detect total RNA in a sample without differentiating vRNA from +RNA. These assays are not designed to distinguish IAV infection versus exposure of an individual to an environment enriched with IAVs but wherein no viral replication occurs. We therefore developed a strand-specific hybridization (SSH) assay that differentiates between vRNA and +RNA and quantifies relative levels of each RNA species.

Funder

HHS | Centers for Disease Control and Prevention

Publisher

American Society for Microbiology

Subject

Microbiology (medical)

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