Validation of the Seegene RV15 multiplex PCR for the detection of influenza A subtypes and influenza B lineages during national influenza surveillance in hospitalized adults

J J LeBlanc¹, M ElSherif¹, S Mulpuru², M Warhuus¹, A Ambrose¹, M Andrew¹, G Boivin³, W Bowie⁴, A Chit^{5

6}, G Dos Santos^{7

8}, K Green⁹, S A Halperin¹, T F Hatchette¹, B Ibarguchi^{10

11}, J Johnstone¹², K Katz¹³, J M Langley¹, P Lagacé-Wiens¹⁴, M Loeb¹², A Lund¹, D MacKinnon-Cameron¹, A McCarthy¹⁵, J E McElhaney¹⁶, A McGeer⁹, A Poirier¹⁷, J Powis¹⁸, D Richardson¹⁹, M Semret²⁰, V Shinde^{21

22}, D Smyth²³, S Trottier³, L Valiquette²⁴, D Webster²⁵, L Ye¹, S A McNeil²

Affiliations

¹ Canadian Center for Vaccinology, Dalhousie University, IWK Health Centre, and Nova Scotia Health Authority, Halifax, NS, Canada.
² Ottawa Hospital Research Institute, University of Ottawa, Ottawa, ON, Canada.
³ Centre Hospitalier Universitaire de Québec, QC, Canada.
⁴ University of British Columbia, Vancouver, BC, Canada.
⁵ Sanofi Pasteur, Swiftwater, PA, USA.
⁶ Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada.
⁷ Present address: GSK, Wavre, Belgium.
⁸ Business & Decision Life Sciences (on behalf of GSK), Bruxelles, Belgium.
⁹ Mount Sinai Hospital, Toronto, ON, Canada.
¹⁰ Present address: Bayer, Inc., Mississauga, Ontario, Canada.
¹¹ GSK, Mississauga, ON, Canada.
¹² Public Health Ontario and University of Toronto, Toronto, ON, Canada.
¹³ North York General Hospital, Toronto, ON, Canada.
¹⁴ St Boniface Hospital, Winnipeg, MB, Canada.
¹⁵ Ottawa Hospital General, Ottawa, Ontario, Canada.
¹⁶ Health Sciences North Research Institute, Sudbury, ON, Canada.
¹⁷ Centre Intégré Universitaire de Santé et Services Sociaux, Quebec, QC, Canada.
¹⁸ Toronto East General Hospital, Toronto, ON, Canada.
¹⁹ William Osler Health System, Brampton, ON, Canada.
²⁰ McGill University, Montreal, QC, Canada.
²¹ GSK, King of Prussia, PA, USA.
²² Present address: Novavax Vaccines, Washington, DC, USA.
²³ The Moncton Hospital, Moncton, NB, Canada.
²⁴ Université de Sherbrooke, Sherbrooke, QC, Canada.
²⁵ Horizon Health, Saint John, NB, Canada.

PMID: 31264957
PMCID: PMC7431100
DOI: 10.1099/jmm.0.001032

Validation of the Seegene RV15 multiplex PCR for the detection of influenza A subtypes and influenza B lineages during national influenza surveillance in hospitalized adults

J J LeBlanc et al. J Med Microbiol. 2020 Feb.

. 2020 Feb;69(2):256-264.

doi: 10.1099/jmm.0.001032. Epub 2019 Jun 28.

Authors

Affiliations

¹ Canadian Center for Vaccinology, Dalhousie University, IWK Health Centre, and Nova Scotia Health Authority, Halifax, NS, Canada.
² Ottawa Hospital Research Institute, University of Ottawa, Ottawa, ON, Canada.
³ Centre Hospitalier Universitaire de Québec, QC, Canada.
⁴ University of British Columbia, Vancouver, BC, Canada.
⁵ Sanofi Pasteur, Swiftwater, PA, USA.
⁶ Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada.
⁷ Present address: GSK, Wavre, Belgium.
⁸ Business & Decision Life Sciences (on behalf of GSK), Bruxelles, Belgium.
⁹ Mount Sinai Hospital, Toronto, ON, Canada.
¹⁰ Present address: Bayer, Inc., Mississauga, Ontario, Canada.
¹¹ GSK, Mississauga, ON, Canada.
¹² Public Health Ontario and University of Toronto, Toronto, ON, Canada.
¹³ North York General Hospital, Toronto, ON, Canada.
¹⁴ St Boniface Hospital, Winnipeg, MB, Canada.
¹⁵ Ottawa Hospital General, Ottawa, Ontario, Canada.
¹⁶ Health Sciences North Research Institute, Sudbury, ON, Canada.
¹⁷ Centre Intégré Universitaire de Santé et Services Sociaux, Quebec, QC, Canada.
¹⁸ Toronto East General Hospital, Toronto, ON, Canada.
¹⁹ William Osler Health System, Brampton, ON, Canada.
²⁰ McGill University, Montreal, QC, Canada.
²¹ GSK, King of Prussia, PA, USA.
²² Present address: Novavax Vaccines, Washington, DC, USA.
²³ The Moncton Hospital, Moncton, NB, Canada.
²⁴ Université de Sherbrooke, Sherbrooke, QC, Canada.
²⁵ Horizon Health, Saint John, NB, Canada.

PMID: 31264957
PMCID: PMC7431100
DOI: 10.1099/jmm.0.001032

Abstract

Background. The Serious Outcomes Surveillance Network of the Canadian Immunization Research Network (CIRN SOS) has been performing active influenza surveillance since 2009 (ClinicalTrials.gov identifier: NCT01517191). Influenza A and B viruses are identified and characterized using real-time reverse-transcriptase polymerase chain reaction (RT-PCR), and multiplex testing has been performed on a subset of patients to identify other respiratory virus aetiologies. Since both methods can identify influenza A and B, a direct comparison was performed.Methods. Validated real-time RT-PCRs from the World Health Organization (WHO) to identify influenza A and B viruses, characterize influenza A viruses into the H1N1 or H3N2 subtypes and describe influenza B viruses belonging to the Yamagata or Victoria lineages. In a subset of patients, the Seeplex RV15 One-Step ACE Detection assay (RV15) kit was also used for the detection of other respiratory viruses.Results. In total, 1111 nasopharyngeal swabs were tested by RV15 and real-time RT-PCRs for influenza A and B identification and characterization. For influenza A, RV15 showed 98.0 % sensitivity, 100 % specificity and 99.7 % accuracy. The performance characteristics of RV15 were similar for influenza A subtypes H1N1 and H3N2. For influenza B, RV15 had 99.2 % sensitivity, 100 % specificity and 99.8 % accuracy, with similar assay performance being shown for both the Yamagata and Victoria lineages.Conclusions. Overall, the detection of circulating subtypes of influenza A and lineages of influenza B by RV15 was similar to detection by real-time RT-PCR. Multiplex testing with RV15 allows for a more comprehensive respiratory virus surveillance in hospitalized adults, without significantly compromising the reliability of influenza A or B virus detection.

Keywords: influenza; lineage; multiplex PCR; subtype; validation.

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Conflict of interest statement

J. L. received research grants from Merck for work outside the study, but no personal payments. T. F. H. and S. A. M. report payments to their institution from the GSK group of companies for the conduct of the study, and payments from Pfizer, Merck, Novartis and Sanofi‐Pasteur outside the submitted work. M. K. A. reports grants from the GSK group of companies, Pfizer and Sanofi, but no personal payments. J. M. reports payments to her institution from the GSK group of companies and Sanofi for her participation in advisory boards. A. P. reports payments from Actelion, Sanofi‐Pasteur and Genentech. J. P. reports payments from the GSK group of companies, Merck, Roche and Synthetic Biologics outside the submitted work. L. V. received research grants from the GSK group of companies, Pfizer, Optimer, Cubist and Merck, and personal fees from Merck, Optimer and Cubist. G. D. S. reports he was external consultant at Business and Decision Life Sciences (on behalf of GSK) at the time of the study, and is currently employed by the GSK group of companies and holds shares in the GSK group of companies. No other conflicts were declared.

Figures

**Fig. 1.**
Overlap of positive influenza A or B results between RV15 and the WHO real-time RT-PCRs. Venn diagrams illustrating the overlap of positive results for influenza A or influenza B in patients tested by the Seegene multiplex PCR (RV15), the WHO real-time RT-PCRs for influenza A or B detection (A/B) and influenza A subtyping (H1/H3) or influenza B discrimination for Yamagata or Victoria (Yam/Vic) lineages.

**Fig. 2.**
Overlap of results between RV15 and the WHO real-time RT-PCRs for characterized influenza A or B viruses. Venn diagram illustrating the overlap of positive results between RV15 and influenza A/B real-time RT-PCRs for characterized influenza A subtypes (H1N1 or H3N2) and influenza B lineages (Yamagata or Victoria).

See this image and copyright information in PMC

References

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Validation of the Seegene RV15 multiplex PCR for the detection of influenza A subtypes and influenza B lineages during national influenza surveillance in hospitalized adults

Affiliations

Validation of the Seegene RV15 multiplex PCR for the detection of influenza A subtypes and influenza B lineages during national influenza surveillance in hospitalized adults

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

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Associated data

LinkOut - more resources

Full Text Sources

Medical

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