Multiplex Assay for Simultaneously Typing and Subtyping Influenza Viruses by Use of an Electronic Microarray
Ying Huang,Huong Tang,Stuart Duffy,Yuwen Hong,Sylvia A. Norman,Madhu Ghosh,Jie He,Michael E. Bose,Kelly J. Henrickson,Jiang Fan,Andrea J. Kraft,William G. Weisburg,Elizabeth L. Mather +12 more
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TLDR
The assay is a rapid, accurate, user-friendly method for simultaneously typing and subtyping influenza viruses and to distinguish influenza A virus subtypes H1N1 and H3N2 from the potentially pandemic avian virus subtype H5N1.Abstract:Â
We report on the use of an electronic microarray to simultaneously type influenza A and B viruses and to distinguish influenza A virus subtypes H1N1 and H3N2 from the potentially pandemic avian virus subtype H5N1. The assay targets seven genes: the H1, H3, H5, N1, and N2 genes of influenza A virus; the matrix protein M1 gene of influenza A virus; and the nonstructural protein (NS) gene of influenza B virus. By combining a two-step reverse transcription-multiplex PCR with typing and subtyping on the electronic microarray, the assay achieved an analytical sensitivity of 102 to 103 copies of transcripts per reaction for each of the genes. The assay correctly typed and subtyped 15 different influenza virus isolates, including two influenza B virus, five A/H1N1, six A/H3N2, and two A/H5N1 isolates. In addition, the assay correctly identified 8 out of 10 diluted, archived avian influenza virus specimens with complete typing and subtyping information and 2 specimens with partial subtyping information. In a study of 146 human clinical specimens that had previously been shown to be positive for influenza virus or another respiratory virus, the assay showed a clinical sensitivity of 96% and a clinical specificity of 100%. The assay is a rapid, accurate, user-friendly method for simultaneously typing and subtyping influenza viruses.read more
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Journal ArticleDOI
Update on Influenza Diagnostics: Lessons from the Novel H1N1 Influenza A Pandemic
TL;DR: The need and role for subtyping of influenza viruses and antiviral susceptibility testing will likely depend on qualitative (circulating subtypes and their resistance patterns) and quantitative (relative prevalence) characterization of influenza virus circulating during future epidemics and pandemics.
Journal ArticleDOI
Methods for molecular surveillance of influenza.
TL;DR: The application of these techniques, including reverse transcriptase-PCR, real-time PCR, microarrays and other nucleic acid sequencing-based amplifications have greatly enhanced the capability for surveillance and characterization of influenza viruses.
Journal ArticleDOI
Rapid Multiplex Reverse Transcription-PCR Typing of Influenza A and B Virus, and Subtyping of Influenza A Virus into H1, 2, 3, 5, 7, 9, N1 (Human), N1 (Animal), N2, and N7, Including Typing of Novel Swine Origin Influenza A (H1N1) Virus, during the 2009 Outbreak in Milwaukee, Wisconsin
Jie He,Michael E. Bose,Eric T. Beck,Jiang Fan,Sagarika Tiwari,Jacob A. Metallo,Lisa A. Jurgens,Sue C. Kehl,Nathan A. Ledeboer,Swati Kumar,William G. Weisburg,Kelly J. Henrickson +11 more
TL;DR: FluPlex as mentioned in this paper is a reverse transcription-PCR enzyme hybridization (RTP-EHP) based method for the detection and subtyping of influenza viruses, which was used to confirm the diagnoses for the first infected patients in Wisconsin.
Journal ArticleDOI
Molecular diagnosis of respiratory viruses
Eric T. Beck,Kelly J. Henrickson +1 more
TL;DR: Molecular detection has many proven advantages over standard virological methods and will further separate itself through increased multiplexing, processing speed and automation.
Journal ArticleDOI
Development of a Rapid Automated Influenza A, Influenza B, and Respiratory Syncytial Virus A/B Multiplex Real-Time RT-PCR Assay and Its Use during the 2009 H1N1 Swine-Origin Influenza Virus Epidemic in Milwaukee, Wisconsin
Eric T. Beck,Lisa A. Jurgens,Sue C. Kehl,Sue C. Kehl,Michael E. Bose,Teresa N. Patitucci,Elizabeth LaGue,Patrick Darga,Kimberly Wilkinson,Lorraine M. Witt,Jiang Fan,Jie He,Swati Kumar,Swati Kumar,Kelly J. Henrickson,Kelly J. Henrickson +15 more
TL;DR: Rapid, semiautomated, and fully automated multiplex real-time RT-PCR assays were developed and validated for the detection of influenza (Flu) A, Flu B, and respiratory syncytial virus (RSV) from nasopharyngeal specimens and were effectively used to distinguish Flu A infections from Flu B and RSV infections during the current S-OIV outbreak in Milwaukee, WI.
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