1. Single assay for simultaneous detection and differential identification of human and avian influenza virus types, subtypes, and emergent variants.
- Author
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Metzgar D, Myers CA, Russell KL, Faix D, Blair PJ, Brown J, Vo S, Swayne DE, Thomas C, Stenger DA, Lin B, Malanoski AP, Wang Z, Blaney KM, Long NC, Schnur JM, Saad MD, Borsuk LA, Lichanska AM, Lorence MC, Weslowski B, Schafer KO, and Tibbetts C
- Subjects
- Animals, Base Sequence, Birds, Cell Line, Chick Embryo, Gene Expression Profiling methods, Gene Expression Regulation, Viral, Humans, Influenza A virus classification, Influenza in Birds virology, Influenza, Human virology, Molecular Sequence Data, Oligonucleotide Array Sequence Analysis, Reproducibility of Results, Reverse Transcriptase Polymerase Chain Reaction, Sensitivity and Specificity, Sequence Analysis, DNA, Sequence Homology, Nucleic Acid, Influenza A virus genetics, Influenza in Birds diagnosis, Influenza, Human diagnosis, Pathology, Molecular methods
- Abstract
For more than four decades the cause of most type A influenza virus infections of humans has been attributed to only two viral subtypes, A/H1N1 or A/H3N2. In contrast, avian and other vertebrate species are a reservoir of type A influenza virus genome diversity, hosting strains representing at least 120 of 144 combinations of 16 viral hemagglutinin and 9 viral neuraminidase subtypes. Viral genome segment reassortments and mutations emerging within this reservoir may spawn new influenza virus strains as imminent epidemic or pandemic threats to human health and poultry production. Traditional methods to detect and differentiate influenza virus subtypes are either time-consuming and labor-intensive (culture-based) or remarkably insensitive (antibody-based). Molecular diagnostic assays based upon reverse transcriptase-polymerase chain reaction (RT-PCR) have short assay cycle time, and high analytical sensitivity and specificity. However, none of these diagnostic tests determine viral gene nucleotide sequences to distinguish strains and variants of a detected pathogen from one specimen to the next. Decision-quality, strain- and variant-specific pathogen gene sequence information may be critical for public health, infection control, surveillance, epidemiology, or medical/veterinary treatment planning. The Resequencing Pathogen Microarray (RPM-Flu) is a robust, highly multiplexed and target gene sequencing-based alternative to both traditional culture- or biomarker-based diagnostic tests. RPM-Flu is a single, simultaneous differential diagnostic assay for all subtype combinations of type A influenza viruses and for 30 other viral and bacterial pathogens that may cause influenza-like illness. These other pathogen targets of RPM-Flu may co-infect and compound the morbidity and/or mortality of patients with influenza. The informative specificity of a single RPM-Flu test represents specimen-specific viral gene sequences as determinants of virus type, A/HN subtype, virulence, host-range, and resistance to antiviral agents.
- Published
- 2010
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