Rapid Semiautomated Subtyping of Influenza Virus Species during the 2009 Swine Origin Influenza A H1N1 Virus Epidemic in Milwaukee, Wisconsin

• Published in: Journal of Clinical Microbiology 2009-09, Vol.47 (9), p.2779-2786
• Main Authors: Bose, Michael E, Beck, Eric T, Ledeboer, Nate, Kehl, Sue C, Jurgens, Lisa A, Patitucci, Teresa, Witt, Lorraine, LaGue, Elizabeth, Darga, Patrick, He, Jie, Fan, Jiang, Kumar, Swati, Henrickson, Kelly J
• Format: Article
• Language: English
• Subjects: Automation; Biological and medical sciences; Disease Outbreaks; DNA Primers - genetics; Fundamental and applied biological sciences. Psychology; Genotype; Humans; Influenza A virus; Influenza A virus - classification; Influenza A virus - genetics; Influenza A virus - isolation & purification; Influenza A Virus, H1N1 Subtype - isolation & purification; Influenza B virus; Influenza B virus - genetics; Influenza B virus - isolation & purification; Influenza, Human - epidemiology; Influenza, Human - virology; Microbiology; Miscellaneous; Reproducibility of Results; Respiratory syncytial virus; Respiratory Syncytial Viruses - isolation & purification; Sensitivity and Specificity; Virology; Virology - methods; Wisconsin - epidemiology
• ISSN: 0095-1137; 1098-660X
• DOI: 10.1128/JCM.00999-09

In the spring of 2009, a novel influenza A (H1N1) virus (swine origin influenza virus [S-OIV]) emerged and began causing a large outbreak of illness in Milwaukee, WI. Our group at the Midwest Respiratory Virus Program laboratory developed a semiautomated real-time multiplex reverse transcription-PCR assay (Seasonal), employing the NucliSENS easyMAG system (bioMérieux, Durham, NC) and a Raider thermocycler (HandyLab Inc., Ann Arbor, MI), that typed influenza A virus, influenza B virus, and respiratory syncytial virus (RSV) and subtyped influenza A virus into the currently circulating H1 and H3 subtypes, as well as a similar assay that identified H1 of S-OIV. The Seasonal and H1 S-OIV assays demonstrated analytical limits of detection of