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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.

  • LeBlanc, J J1
  • ElSherif, M1
  • Mulpuru, S2
  • Warhuus, M1
  • Ambrose, A1
  • Andrew, M1
  • Boivin, G3
  • Bowie, W4
  • Chit, A5, 6
  • Dos Santos, G7, 8
  • Green, K9
  • Halperin, S A1
  • Hatchette, T F1
  • Ibarguchi, B10, 11
  • Johnstone, J12
  • Katz, K13
  • Langley, J M1
  • Lagacé-Wiens, P14
  • Loeb, M12
  • Lund, A1
  • And 15 more
  • 1 Canadian Center for Vaccinology, Dalhousie University, IWK Health Centre, and Nova Scotia Health Authority, Halifax, NS, Canada. , (Canada)
  • 2 Ottawa Hospital Research Institute, University of Ottawa, Ottawa, ON, Canada. , (Canada)
  • 3 Centre Hospitalier Universitaire de Québec, QC, Canada. , (Canada)
  • 4 University of British Columbia, Vancouver, BC, Canada. , (Canada)
  • 5 Sanofi Pasteur, Swiftwater, PA, USA.
  • 6 Leslie Dan Faculty of Pharmacy, University of Toronto, Toronto, ON, Canada. , (Canada)
  • 7 Business & Decision Life Sciences (on behalf of GSK), Bruxelles, Belgium. , (Belgium)
  • 8 Present address: GSK, Wavre, Belgium. , (Belgium)
  • 9 Mount Sinai Hospital, Toronto, ON, Canada. , (Canada)
  • 10 GSK, Mississauga, ON, Canada. , (Canada)
  • 11 Present address: Bayer, Inc., Mississauga, Ontario, Canada. , (Canada)
  • 12 Public Health Ontario and University of Toronto, Toronto, ON, Canada. , (Canada)
  • 13 North York General Hospital, Toronto, ON, Canada. , (Canada)
  • 14 St Boniface Hospital, Winnipeg, MB, Canada. , (Canada)
  • 15 Ottawa Hospital General, Ottawa, Ontario, Canada. , (Canada)
  • 16 Health Sciences North Research Institute, Sudbury, ON, Canada. , (Canada)
  • 17 Centre Intégré Universitaire de Santé et Services Sociaux, Quebec, QC, Canada. , (Canada)
  • 18 Toronto East General Hospital, Toronto, ON, Canada. , (Canada)
  • 19 William Osler Health System, Brampton, ON, Canada. , (Canada)
  • 20 McGill University, Montreal, QC, Canada. , (Canada)
  • 21 GSK, King of Prussia, PA, USA.
  • 22 Present address: Novavax Vaccines, Washington, DC, USA.
  • 23 The Moncton Hospital, Moncton, NB, Canada. , (Canada)
  • 24 Université de Sherbrooke, Sherbrooke, QC, Canada. , (Canada)
  • 25 Horizon Health, Saint John, NB, Canada. , (Canada)
Published Article
Journal of Medical Microbiology
Microbiology Society
Publication Date
Feb 01, 2020
DOI: 10.1099/jmm.0.001032
PMID: 31264957


Background. The Serious Outcomes Surveillance Network of the Canadian Immunization Research Network (CIRN SOS) has been performing active influenza surveillance since 2009 ( 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.

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