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Volume 16
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10.3390/v16101557
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Discriminating North American Swine Influenza Viruses with a Portable, One-Step, Triplex Real-Time RT-PCR Assay, and Portable Sequencing

by
Marie K. Kirby
1,†,
Bo Shu
1,†,
Matthew W. Keller
1,
Malania M. Wilson
1,
Benjamin L. Rambo-Martin
1,
Yunho Jang
1,
Jimma Liddell
1,
Eduardo Salinas Duron
1,2,
Jacqueline M. Nolting
3,
Andrew S. Bowman
3,
C. Todd Davis
1,
David E. Wentworth
1,‡ and
John R. Barnes
1,*
1
Virology, Surveillance and Diagnostic Branch, Influenza Division, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA
2
Chippewa Government Solutions (CGS), Sault Tribe Incorporated (STI), Cherokee Nation Operational Solutions (CNOS), Sault Sainte Marie, MI 49783, USA
3
Department of Veterinary Preventive Medicine, The Ohio State University, Columbus, OH 43210, USA
*
Author to whom correspondence should be addressed.
†
These authors contributed equally to this work.
‡
Current address: CORVD Division, Centers for Disease Control and Prevention, Atlanta, GA 30329, USA.
Viruses 2024, 16(10), 1557; https://doi.org/10.3390/v16101557
Submission received: 10 September 2024 / Revised: 27 September 2024 / Accepted: 28 September 2024 / Published: 30 September 2024
(This article belongs to the Special Issue Advances in Animal Influenza Virus Research: Third Edition)
Review Reports Versions Notes

Abstract

Swine harbors a genetically diverse population of swine influenza A viruses (IAV-S), with demonstrated potential to transmit to the human population, causing outbreaks and pandemics. Here, we describe the development of a one-step, triplex real-time reverse transcription-polymerase chain reaction (rRT-PCR) assay that detects and distinguishes the majority of the antigenically distinct influenza A virus hemagglutinin (HA) clades currently circulating in North American swine, including the IAV-S H1 1A.1 (α), 1A.2 (β), 1A.3 (γ), 1B.2.2 (δ1) and 1B.2.1 (δ2) clades, and the IAV-S H3 2010.1 clade. We performed an in-field test at an exhibition swine show using in-field viral concentration and RNA extraction methodologies and a portable real-time PCR instrument, and rapidly identified three distinct IAV-S clades circulating within the N.A. swine population. Portable sequencing is used to further confirm the results of the in-field test of the swine triplex assay. The IAV-S triplex rRT-PCR assay can be easily transported and used in-field to characterize circulating IAV-S clades in North America, allowing for surveillance and early detection of North American IAV-S with human outbreak and pandemic potential.
Keywords: swine influenza virus; triplex real-time RT-PCR; portable sequencing swine influenza virus; triplex real-time RT-PCR; portable sequencing

Share and Cite

MDPI and ACS Style

Kirby, M.K.; Shu, B.; Keller, M.W.; Wilson, M.M.; Rambo-Martin, B.L.; Jang, Y.; Liddell, J.; Salinas Duron, E.; Nolting, J.M.; Bowman, A.S.; et al. Discriminating North American Swine Influenza Viruses with a Portable, One-Step, Triplex Real-Time RT-PCR Assay, and Portable Sequencing. Viruses 2024, 16, 1557. https://doi.org/10.3390/v16101557

AMA Style

Kirby MK, Shu B, Keller MW, Wilson MM, Rambo-Martin BL, Jang Y, Liddell J, Salinas Duron E, Nolting JM, Bowman AS, et al. Discriminating North American Swine Influenza Viruses with a Portable, One-Step, Triplex Real-Time RT-PCR Assay, and Portable Sequencing. Viruses. 2024; 16(10):1557. https://doi.org/10.3390/v16101557

Chicago/Turabian Style

Kirby, Marie K., Bo Shu, Matthew W. Keller, Malania M. Wilson, Benjamin L. Rambo-Martin, Yunho Jang, Jimma Liddell, Eduardo Salinas Duron, Jacqueline M. Nolting, Andrew S. Bowman, and et al. 2024. "Discriminating North American Swine Influenza Viruses with a Portable, One-Step, Triplex Real-Time RT-PCR Assay, and Portable Sequencing" Viruses 16, no. 10: 1557. https://doi.org/10.3390/v16101557

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