Development and evaluation of a real‐time nucleic acid sequence based amplification assay for rapid detection of influenza A
Identifieur interne : 005D57 ( Main/Merge ); précédent : 005D56; suivant : 005D58Development and evaluation of a real‐time nucleic acid sequence based amplification assay for rapid detection of influenza A
Auteurs : Catherine Moore [Royaume-Uni] ; Sam Hibbitts [Royaume-Uni] ; Neil Owen [Royaume-Uni] ; Sally A. Corden [Royaume-Uni] ; Graham Harrison [Royaume-Uni] ; Julie Fox [Canada] ; Colin Gelder [Royaume-Uni] ; Diana Westmoreland [Royaume-Uni]Source :
- Journal of Medical Virology [ 0146-6615 ] ; 2004-12.
English descriptors
- Teeft :
- Antibody titer, Assay, Assay evaluation, Beacon, Catherine moore, Cell culture, Cell suspension, Clin, Clinical excellence, Clinical samples, Community populations, Convalescent, Convalescent sample, Copy number, Detection rate, Elli, Example results, Genetic shift, Hibbitts, Internal control, Kappa statistic, Lysis buffer, Microbiol, Molecular assays, Molecular beacon, Molecular techniques, Monoclonal antibodies, More methods, Nasba, Nasba assay, Nasba assays, Nasba detection, Nasba reaction, Nasba results, Nasba testing, Nasopharyngeal aspirates, National institute, Negative control signal, Negative result, Negative samples, Neuraminidase, Neuraminidase genes, Neuraminidase inhibitors, Nphs microbiology cardiff, Nucleic, Nucleic acid, Nucleic acid sequence, Nursing home, Optical density, Oseltamivir, Oseltamivir treatment, Outbreak, Patient group, Positive result, Positive samples, Post mortem tissue samples, Primary care centers, Primer, Prototype strains, Rapid diagnosis, Reaction volume, Respiratory samples, Respiratory swabs, Respiratory viruses, Sampling rate, Serial dilutions, Serology, Severe illness, Sigmoid curve, Simple method, Single reaction, Swab, Throat swabs, Total population, Traditional laboratory techniques, Traditional methods, University hospital, Uorescence signal, Virol, Virus, Virus input, Virus stock, Wales specialist virology centre, Welsh assembly government, Wide range, Zambon.
Abstract
The development and introduction of effective treatment for influenza A in the form of neuraminidase inhibitors have made the rapid diagnosis of infection important especially in high‐risk populations. The aim of this study was to develop a real‐time nucleic acid sequenced based amplification (NASBA) using a molecular beacon that could detect a wide range of influenza A subtypes and strains in a single reaction by targeting a conserved region of the influenza genome, and to evaluate its sensitivity and specificity against traditional laboratory techniques on a range of clinical samples usefulness during the 2003/2004 influenza season. The results demonstrated the assay to be highly sensitive and specific, detecting <0.1 TCID50 of virus stock. Three hundred eighty‐nine clinical samples were tested in total from two patient groups. Overall, the real‐time NASBA assay detected 64% (66/103) more influenza positive samples than cell culture and direct immunofluorescence (IF) and, therefore, was shown to be more sensitive in detecting influenza A in a wide range of respiratory samples than traditional methods. In conclusion, the real‐time influenza A assay demonstrated clinical usefulness in both hospital and community populations. J. Med. Virol. 74:619–628, 2004. © 2004 Wiley‐Liss, Inc.
Url:
DOI: 10.1002/jmv.20221
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<term>Cell suspension</term>
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<term>University hospital</term>
<term>Uorescence signal</term>
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<term>Virus</term>
<term>Virus input</term>
<term>Virus stock</term>
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<front><div type="abstract" xml:lang="en">The development and introduction of effective treatment for influenza A in the form of neuraminidase inhibitors have made the rapid diagnosis of infection important especially in high‐risk populations. The aim of this study was to develop a real‐time nucleic acid sequenced based amplification (NASBA) using a molecular beacon that could detect a wide range of influenza A subtypes and strains in a single reaction by targeting a conserved region of the influenza genome, and to evaluate its sensitivity and specificity against traditional laboratory techniques on a range of clinical samples usefulness during the 2003/2004 influenza season. The results demonstrated the assay to be highly sensitive and specific, detecting <0.1 TCID50 of virus stock. Three hundred eighty‐nine clinical samples were tested in total from two patient groups. Overall, the real‐time NASBA assay detected 64% (66/103) more influenza positive samples than cell culture and direct immunofluorescence (IF) and, therefore, was shown to be more sensitive in detecting influenza A in a wide range of respiratory samples than traditional methods. In conclusion, the real‐time influenza A assay demonstrated clinical usefulness in both hospital and community populations. J. Med. Virol. 74:619–628, 2004. © 2004 Wiley‐Liss, Inc.</div>
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