Design and clinical application of a molecular method for detection and typing of the influenza A/H1N1pdm virus

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• 作者：Eleonora Lalle ; Licia Bordi ; Concetta Castilletti ; Silvia Meschi ; Marina Selleri ; Fabrizio Carletti ; Daniele Lapa ; Damiano Travaglini ; Giuseppe Ippolito ; Maria Rosaria Capobianchi ; Antonino Di Caro
• 关键词：Influenza A/H1N1pdm ; S-OIV ; Influenza pandemic ; Molecular diagnosis ; RT-PCR ; Sequencing ; Typing
• 刊名：Journal of Virological Methods
• 出版年：2010
• 出版时间：February 2010
• 年：2010
• 卷：163
• 期：2
• 页码：486-488
• 全文大小：319 K

文摘

In March/April 2009, Mexico experienced an outbreak of respiratory illness, due to a new influenza of swine origin virus, which spread rapidly via human-to-human transmission, and became pandemic (A/H1N1pdm). Because of its unique genome composition, which includes gene segments of swine, avian and human origin, and to the considerable differences to the human influenza A viruses that have circulated so far, the currently used molecular methods proved inadequate.

Based on published sequences, a primer set targeting the nucleoprotein gene was designed, which provided enhanced sensitivity for the new strain and proved suitable for sequence-based strain identification. The novel nucleoprotein reverse-transcription-PCR showed higher sensitivity for A/H1N1pdm than a commercial test for influenza A, and was comparable to the real-time-based method developed by the Centers for Disease Control and Prevention. It was used to screen 177 clinical samples referred to the laboratory for suspected A/H1N1pdm infection, detecting 17 (9.6 % ) infections that were confirmed by sequence analysis (100 % sensitivity as compared to the real-time kit).

The novel method is suitable for the diagnosis of A/H1N1pdm, and is also suitable, at least in the screening phase, for laboratories not equipped with the real-time PCR technology.