Development and evaluation of a real-time nucleic acid sequence based amplification assay for rapid detection of influenza A

✍ Scribed by Catherine Moore; Sam Hibbitts; Neil Owen; Sally A. Corden; Graham Harrison; Julie Fox; Colin Gelder; Diana Westmoreland

• Publisher: John Wiley and Sons
• Year: 2004
• Tongue: English
• Weight: 129 KB
• Volume: 74
• Category: Article
• ISSN: 0146-6615
• DOI: 10.1002/jmv.20221

✦ Synopsis

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 TCID~50~ 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.