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

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• 作者：Carla Duncan^a ; ¹ ; Jennifer L. Guthrie^a ; ¹ ; Nathalie Tijet^a ; Naglaa Elgngihy^a ; Christine Turenne^a ; Christine Seah^a ; Rachel Lau^a ; Lisa McTaggart^a ; Gustavo Mallo^a ; Stephen Perusini^a ; Anu Rebbapragada^a ; ^b ; ^c ; Roberto Melano^a ; ^b ; ^c ; Donald E. Low^a ; ^b ; ^c ; David Farrell^a ; ^b ; Cyril Guyard^a ; ^b ; ^c ; ^{cyril.guyard@oahpp.ca}
• 关键词：rRT-PCR ; S-OIV H1N1 FluA ; Design ; Validation
• 刊名：Diagnostic Microbiology and Infectious Disease
• 出版年：2011
• 出版时间：February 2011
• 年：2011
• 卷：69
• 期：2
• 页码：167-171
• 全文大小：184 K

文摘

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.