DTU Nanotech, Department of Micro- and Nanotechnology, Technical University of Denmark, Ørsteds Plads, DK-2800 Kgs. Lyngby, Denmark.
Diagn Microbiol Infect Dis. 2011 Apr;69(4):432-9. doi: 10.1016/j.diagmicrobio.2010.11.008.
Endemic of avian influenza virus (AIV) in Asia and epizootics in some European regions have caused considerable public concern on a possible pandemic of AIV. A rapid method for virus detection and effective surveillance in wild avian, poultry production as well as in humans is required. In this article, a DNA microarray-based solid-phase polymerase chain reaction (PCR) approach has been developed for rapid detection of influenza virus type A and for simultaneous identification of pathogenic virus subtypes H5 and H7. This solid-phase RT-PCR method combined reverse-transcription amplification of RNA extract in the liquid phase with sequence-specific nested PCR on the solid phase. A simple ultraviolet cross-linking method was used to immobilize the DNA probes over an unmodified glass surface, which makes solid-phase PCR a convenient possibility for AIV screening. The testing of 33 avian fecal and tracheal swab specimens was completed in less than 2 h with 94% accuracy.
亚洲禽流感病毒（AIV）的流行以及一些欧洲地区的禽流感疫情引起了公众对 AIV 可能大流行的极大关注。需要一种快速的方法来检测病毒，并对野生鸟类、家禽生产以及人类进行有效的监测。在本文中，我们开发了一种基于 DNA 微阵列的固相聚合酶链反应（PCR）方法，用于快速检测甲型流感病毒，并同时鉴定致病性病毒亚型 H5 和 H7。这种固相 RT-PCR 方法将 RNA 提取物在液相中的逆转录扩增与固相上的序列特异性巢式 PCR 相结合。我们使用简单的紫外线交联方法将 DNA 探针固定在未经修饰的玻璃表面上，这使得固相 PCR 成为 AIV 筛选的一种便捷选择。对 33 份禽粪便和气管拭子标本的检测在不到 2 小时内完成，准确率为 94%。
