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

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.