ArticleJournal of clinical microbiology2007
Design and validation of an H5 TaqMan real-time one-step reverse transcription-PCR and confirmatory assays for diagnosis and verification of influenza A virus H5 infections in humans.
Article in Journal of clinical microbiology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- Advances in Detection Techniques for the H5N1 Avian Influenza Virus.International journal of molecular sciences · 2023Review
- Streptavidin-biotin-based directional double Nanobody sandwich ELISA for clinical rapid and sensitive detection of influenza H5N1.Journal of translational medicine · 2014Article
- Development of a sensitive novel diagnostic kit for the highly pathogenic avian influenza A (H5N1) virus.BMC infectious diseases · 2014Article
- Microarray multiplex assay for the simultaneous detection and discrimination of influenza a and influenza B viruses.Indian journal of microbiology · 2014Article
- Review
- Multiplexed, rapid detection of H5N1 using a PCR-free nanoparticle-based genomic microarray assay.BMC biotechnology · 2010Article
- Methods for molecular surveillance of influenza.Expert review of anti-infective therapy · 2010Review
- A real-time RT-PCR for detection of clade 1 and 2 H5N1 influenza A virus using locked nucleic acid (LNA) TaqMan probes.Virology journal · 2010Article
- Development of real time RT-PCR assays for detection of type A influenza virus and for subtyping of avian H5 and H7 hemagglutinin subtypes.Molecular biotechnology · 2010Article
- Emerging molecular assays for detection and characterization of respiratory viruses.Clinics in laboratory medicine · 2009Review
- Universal detection and identification of avian influenza virus by use of resequencing microarrays.Journal of clinical microbiology · 2009Article
- Molecular quantitation of H9N2 avian influenza virus in various organs of broiler chickens using TaqMan real time PCR.Journal of molecular and genetic medicine : an international journal of biomedical research · 2009Article
- design of multiplexed detection assays for identification of avian influenza a virus subtypes pathogenic to humans by SmartCycler real-time reverse transcription-PCR.Journal of clinical microbiology · 2009Article
- Detection of respiratory viruses by molecular methods.Clinical microbiology reviews · 2008Review
- Predicting the sensitivity and specificity of published real-time PCR assays.Annals of clinical microbiology and antimicrobials · 2008Article
- Development and validation of a one-step real-time PCR assay for simultaneous detection of subtype H5, H7, and H9 avian influenza viruses.Journal of clinical microbiology · 2008Article
- Rapid differentiation of influenza A virus subtypes and genetic screening for virus variants by high-resolution melting analysis.Journal of clinical microbiology · 2008Article
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Increasing diversity among influenza H5N1 viruses has resulted in the need for sensitive and specific diagnostic assays, fully validated for the detection of H5 viruses belonging to all hemagglutinin (HA) clades, particularly the recently circulating H5N1 viruses of clade 2. In this report, the development and validation of a real-time, one-step TaqMan reverse transcription-PCR (RT-PCR) assay specific for the detection of influenza A H5 viruses from clades 1, 1', 2, and 3 is described. The real-time assay for H5 virus was shown to be highly sensitive, detecting H5 virus levels of <1 PFU from each of the HA clades. Specificity of the H5 RT-PCR for influenza A H5 viruses was demonstrated by using influenza A viruses of different subtypes, clinical samples containing influenza A viruses H1N1, H3N2, and H5N1, influenza B viruses, and other respiratory viruses. The usefulness of the inclusion of a distinguishable assay positive control and of confirmatory assays for the laboratory diagnosis and verification of H5 virus infections was demonstrated. A real-time RT-PCR pyrosequencing assay, a restriction enzyme digestion assay, and direct sequencing of the H5 real-time RT-PCR amplicon were validated for the confirmation of H5 detection by the diagnostic real-time assay. The H5 real-time assay was applied to diagnostic testing for suspected cases of influenza A virus H5 infection in the United Kingdom. Influenza A H5 viruses were not detected in the cases analyzed; however, influenza A H3N2 virus was detected in 57% of the suspected cases of H5. The H5 TaqMan real-time RT-PCR and confirmatory assays will be useful tools for the laboratory surveillance and rapid diagnosis of H5 infections in humans.
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