Analytical and clinical validation of novel real-time reverse transcriptase-polymerase chain reaction assays for the clinical detection of swine-origin H1N1 influenza viruses

作者：Duncan Carla; Guthrie Jennifer L; Tijet Nathalie; Elgngihy Naglaa; Turenne Christine; Seah Christine; Lau Rachel; McTaggart Lisa; Mallo Gustavo; Perusini Stephen; Rebbapragada Anu; Melano Roberto; Low Donald E; Farrell David; Guyard Cyril^*

来源：Diagnostic Microbiology and Infectious Disease, 2011, 69(2): 167-171.

DOI：10.1016/j.diagmicrobio.2010.09.020

• rRT-PCR
• S-OIV H1N1 FluA
• Design
• Validation

During the early stages of the 2009/2010 swine-origin H1N1 influenza A (S-OIV H1N1 FluA) outbreak, the development and validation of sensitive and specific detection methods were a priority for rapid and accurate diagnosis. Between May and June 2009, 2 real-time reverse transcriptase-polymerase chain reaction (rRT-PCR) assays targeting the hemagglutinin and neuraminidase genes of the S-OIV H1N1 FluA virus were developed. These assays are highly specific, showing no cross-reactivity against a panel of respiratory viruses and can differentiate S-OIV H1N1 from seasonal FluA viruses. Analytical sensitivities of the 2 assays were found to be 10(-1) tissue culture infectious dose, 50%/ml. Clinical testing showed 99.2% sensitivity and 94.6-98.1% specificity. A large prospective analysis showed that 94.8-95.5% of S-OIV positive specimens were negative by seasonal H1/H3 subtyping. The large-scale validation data presented in this report indicate that these novel assays provide an accurate and efficient method for the rapid detection of S-OIV H1N1 FluA viruses.

• 出版日期2011-2