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

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 deve...

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Bibliographic Details
Published in:Journal of Clinical Microbiology 2007-05, Vol.45 (5), p.1535-1543
Main Authors: Ellis, Joanna S, Smith, Joanne W, Braham, Sharleen, Lock, Matthew, Barlow, Katrina, Zambon, Maria C
Format: Article
Language:English
Subjects:
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
Hemagglutinin Glycoproteins, Influenza Virus - isolation & purification
Humans
Infectious diseases
Influenza A virus
Influenza A Virus, H5N1 Subtype - genetics
Influenza A Virus, H5N1 Subtype - isolation & purification
Influenza, Human - diagnosis
Medical sciences
Microbiology
Miscellaneous
Phylogeny
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction - methods
Sensitivity and Specificity
Time Factors
Virology
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Summary: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
ISSN:0095-1137
1098-660X
1098-5530
DOI:10.1128/JCM.02007-06