Faculty Collaboration Database

Rapid semiautomated subtyping of influenza virus species during the 2009 swine origin influenza A H1N1 virus epidemic in Milwaukee, Wisconsin. J Clin Microbiol 2009 Sep;47(9):2779-86

Date

07/31/2009

Pubmed ID

19641066

Pubmed Central ID

PMC2738075

DOI

10.1128/JCM.00999-09

Scopus ID

2-s2.0-69949100084 (requires institutional sign-in at Scopus site)   32 Citations

Abstract

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 <50 50% tissue culture infective doses/ml and 3 to 30 input copies, respectively. Testing of the analytical specificities revealed no cross-reactivity with 41 and 26 different common organisms and demonstrated outstanding reproducibility of results. Clinical testing showed 95% sensitivity for influenza A virus and influenza B virus and 95 and 97% specificity compared to tissue culture. Comparisons of results from other molecular tests showed levels of positive agreement with the Seasonal and H1 S-OIV assay results of 99 and 100% and levels of negative agreement of 98 and 100%. This study has demonstrated the use of a semiautomated system for sensitive, specific, and rapid detection of influenza A virus, influenza B virus, and RSV and subtyping of influenza A virus into human H1 and H3 and S-OIV strains. This assay/system performed well in clinical testing of regular seasonal influenza virus subtypes and was outstanding during the 2009 Milwaukee S-OIV infection outbreak. This recent outbreak of infection with a novel influenza A (H1N1) virus also demonstrates the importance of quickly distributing information on new agents and of having rapid influenza virus subtyping assays widely available for clinical and public health decisions.

Author List

Bose ME, Beck ET, Ledeboer N, Kehl SC, Jurgens LA, Patitucci T, Witt L, LaGue E, Darga P, He J, Fan J, Kumar S, Henrickson KJ

Authors

Michael Bose Research Scientist I in the Pediatrics department at Medical College of Wisconsin
Kelly J. Henrickson MD Professor in the Pediatrics department at Medical College of Wisconsin
Nathan A. Ledeboer PhD Vice Chair, Professor in the Pathology department at Medical College of Wisconsin
Teresa Patitucci PhD Associate Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of Wisconsin

MESH terms used to index this publication - Major topics in bold

Automation
DNA Primers
Disease Outbreaks
Genotype
Humans
Influenza A Virus, H1N1 Subtype
Influenza A virus
Influenza B virus
Influenza, Human
Reproducibility of Results
Respiratory Syncytial Viruses
Sensitivity and Specificity
Virology
Wisconsin