Rapid detection of avian influenza virus in chicken fecal samples by immunomagnetic capture reverse transcriptase-polymerase chain reaction assay
Rapid detection of avian influenza virus in chicken fecal samples by immunomagnetic capture reverse transcriptase-polymerase chain reaction assay
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DOI：
10.1016/j.diagmicrobio.2010.09.022
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发表时间：
2011-03-01
影响因子：
2.9
通讯作者：
Bang, Dang Duong
中科院分区：
文献类型：
--
作者：
Dhumpa, Raghuram;Handberg, Kurt Jensen;Bang, Dang Duong
禽流感病毒（AIV）在世界范围内给家禽业造成巨大的经济损失，并威胁着人类的大流行。禽流感病毒的常规检测方法包括从原始样品如鸟粪中提取病毒RNA并纯化的样品制备。本研究应用磁性微球免疫分离技术从加标的鸡粪便样品中分离纯化禽流感病毒。将珠与抗AIV核蛋白的单克隆抗体缀合，所述AIV核蛋白在所有AIV中是保守的。由于RT-PCR抑制剂的有效去除，因此通过逆转录聚合酶链反应（RT-PCR）检测珠捕获的病毒，而无需RNA提取。所开发的基于珠粒的测定显示出与RNA提取和经典病毒分离方法相当的类似检测限。使用即用型抗体偶联微珠，该方法所需时间少于5小时。此外，该方法有可能集成到芯片实验室系统中，用于快速检测和鉴定AIV。皇冠版权所有（C）2011由爱思唯尔公司出版。All rights reserved.
Avian influenza virus (AIV) causes great economic losses for the poultry industry worldwide and threatens the human population with a pandemic. The conventional detection method for AIV involves sample preparation of viral RNA extraction and purification from raw sample such as bird droppings. In this study, magnetic beads were applied for immunoseparation and purification of AIV from spiked chicken fecal sample. The beads were conjugated with monoclonal antibodies against the AIV nucleoprotein, which is conserved in all the AIV. The bead-captured virus was detected by reverse transcriptase polymerase chain reaction (RT-PCR) without RNA extraction because of effective removal of RT-PCR inhibitors. The developed bead-based assay showed a similar detection limit comparable to the RNA extraction and the classic virus isolation method. Using ready-to-use antibody-conjugated bead, the method requires less than 5 h. Furthermore, the method has potential to integrate into a Lab-on-a-chip system for rapid detection and identification of AIV. Crown Copyright (C) 2011 Published by Elsevier Inc. All rights reserved.