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; M ElSherif; S Mulpuru; M Warhuus; A Ambrose; M Andrew; G Boivin; W Bowie; A Chit; G Dos Santos; K Green; S A Halperin; T F Hatchette; B Ibarguchi; J Johnstone; K Katz; J M Langley; P Lagacé-Wiens; M Loeb; A Lund; D MacKinnon-Cameron; A McCarthy; J E McElhaney; A McGeer; A Poirier; J Powis; D Richardson; M Semret; V Shinde; D Smyth; S Trottier; L Valiquette; D Webster; L Ye; S A McNeil

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 Med Microbiol. 2020 Feb;69(2):256-264. doi: 10.1099/jmm.0.001032. Epub 2019 Jun 28.

Authors

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.

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.

Publication types

Validation Study

MeSH terms

Adult
Canada / epidemiology
Female
Hospitalization
Humans
Influenza A virus / classification
Influenza A virus / genetics
Influenza A virus / isolation & purification*
Influenza B virus / classification
Influenza B virus / genetics
Influenza B virus / isolation & purification*
Influenza, Human / diagnosis
Influenza, Human / epidemiology
Influenza, Human / therapy
Influenza, Human / virology*
Male
Middle Aged
Molecular Diagnostic Techniques / methods*
Multiplex Polymerase Chain Reaction / methods*
Sensitivity and Specificity

Associated data

ClinicalTrials.gov/NCT01517191