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采用磁珠微系统从鸡粪便样本中快速制备用于检测和鉴定禽流感病毒的样品。

Rapid sample preparation for detection and identification of avian influenza virus from chicken faecal samples using magnetic bead microsystem.

机构信息

Laboratory of Applied Micro and Nanotechnology (LAMINATE), National Veterinary Institute (VET), Technical University of Denmark (DTU), Hangøvej 2, DK-8200 Aarhus N, Denmark.

出版信息

J Virol Methods. 2010 Oct;169(1):228-31. doi: 10.1016/j.jviromet.2010.07.017. Epub 2010 Jul 27.

Abstract

Avian influenza virus (AIV) is an infectious agent of birds and mammals. AIV is causing huge economic loss and can be a threat to human health. Reverse transcriptase polymerase chain reaction (RT-PCR) has been used as a method for the detection and identification of AIV virus. Although RT-PCR is a sensitive method for detection of AIV, it requires sample preparation including separation and purification of AIV and concentrate viral RNA. It is laborious and complex process especially for diagnosis using faecal sample. In this study, magnetic beads were used for immunoseparation of AIV in chicken faecal sample by a magnetic microsystem. Using this system, all the 16 hemagglutinin (H) and 9 neuraminidase (N) subtypes of AIV were separated and detected in spiked faecal samples using RT-PCR, without an RNA extraction step. This rapid sample preparation method can be integrated with a total analysis microsystem and used for diagnosis of AIV.

摘要

禽流感病毒(AIV)是一种感染鸟类和哺乳动物的病原体。AIV 会造成巨大的经济损失,并可能对人类健康构成威胁。逆转录聚合酶链反应(RT-PCR)已被用作检测和鉴定 AIV 病毒的方法。尽管 RT-PCR 是一种检测 AIV 的敏感方法,但它需要样本制备,包括 AIV 的分离和纯化以及浓缩病毒 RNA。这是一项繁琐而复杂的过程,特别是对于粪便样本的诊断。在这项研究中,通过磁微系统,使用磁珠对鸡粪便样本中的 AIV 进行免疫分离。使用该系统,无需 RNA 提取步骤,即可通过 RT-PCR 从粪便加标样本中分离和检测到所有 16 种血凝素(H)和 9 种神经氨酸酶(N)亚型的 AIV。这种快速的样本制备方法可以与全分析微系统集成,用于 AIV 的诊断。

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