Incidence of Medically Attended Respiratory Syncytial Virus and Influenza Illnesses in Children 6–59 Months Old During Four Seasons

Simpson, Melissa D.; Kieke, Burney A.; Sundaram, Maria E.; McClure, David L.; Meece, Jennifer K.; Sifakis, Frangiscos; Gasser, Robert A.; Belongia, Edward A.

Sample aliquots were stored at −70°C after initial influenza testing was completed for the influenza vaccine effectiveness studies. Archived samples were tested for the presence of respiratory virus nucleic acid using a multiplex respiratory virus panel (eSensor Respiratory Viral Panel; GenMark Diagnostics, Inc., Carlsbad, CA). This multiplex panel tested for RSV A and B, human rhinovirus, human metapneumovirus, parainfluenza viruses 1-4, influenza A and B (including subtypes of influenza A), coronaviruses OC43, NL63, HKU1, and 229E, and adenoviruses B and E. Nucleic acid was extracted from the swabs using the Roche MagnaPure 2.0 system and was then amplified using RT-PCR with target-specific primers. Target-specific signals were determined by voltammetry, a process that generates electrical signals from ferrocene-labeled signal probes. We validated the GenMark multiplex assay RSV A and B against singleplex assays developed by the Centers for Disease Control and Prevention. The sensitivity and specificity was 98% and 99%, respectively, for RSV (unpublished data, 2012). Other investigators have reported 99% overall agreement between the GenMark multiplex assay and corresponding singleplex RT-PCR for respiratory viruses in children [15] .