Abstract
A real-time NASBA assay for the specific confirmation of influenza A H5N1 infection was developed and evaluated using proficiency panels distributed to the UK influenza network of laboratories and clinical samples received through the Chinese National Influenza Centre in Beijing. The aim of the proficiency panels was to determine the sensitivity and specificity of the assay on a range of influenza virus types and subtypes including different clades of influenza A H5 viruses. The assay was then evaluated using 19 clinical samples obtained from seven confirmed human cases of influenza A H5N1 infection in China. The assay was shown to have a level of sensitivity of 0.01 TCID50 and 10copies/μl using RNA transcripts of the A/VietNam/1194/2004 H5N1 virus. During the evaluation the assay successfully detected H5N1 viruses known to infect humans from clades 1, 2.1, 2.2 and 2.3 as well as low pathogenic H5N3 avian influenza viruses. The clinical utility of the real-time NASBA assay was proven on a range of clinical samples from patients with confirmed H5N1 infection collected during 2005 and 2006. The real-time NASBA assay was demonstrated to be sensitive and rapid allowing for same day confirmation of a H5N1 infection direct from clinical samples.
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