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建立反转录环介导等温扩增检测传染性法氏囊病病毒的方法。

Development of reverse-transcription loop-mediated isothermal amplification for the detection of infectious bursal disease virus.

机构信息

College of Animal Science and Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China.

出版信息

J Virol Methods. 2009 Dec;162(1-2):267-71. doi: 10.1016/j.jviromet.2009.07.010. Epub 2009 Jul 28.

DOI:10.1016/j.jviromet.2009.07.010
PMID:19643144
Abstract

To establish a reverse-transcription loop-mediated isothermal amplification (RT-LAMP) method for rapid detection of infectious bursal disease virus (IBDV), four primers specific to six regions of the VP3 gene were designed; the VP3 region was selected because it is a conserved part of the IBDV genome. After amplification in an isothermal water bath for 70 min, samples containing IBDV generated the expected ladder-like products while other viruses generated no product. The sensitivity and specificity of the RT-LAMP assay were evaluated by comparison with reverse-transcription polymerase chain reaction (RT-PCR) and virus isolation. The assay was significantly more sensitive than normal gel-based RT-PCR. Because it is specific and simple, the RT-LAMP assay can be widely applied in clinical laboratories for rapid detection of IBVD.

摘要

为了建立一种用于快速检测传染性法氏囊病病毒(IBDV)的逆转录环介导等温扩增(RT-LAMP)方法,设计了针对 VP3 基因 6 个区域的 4 个引物;选择 VP3 区域是因为它是 IBDV 基因组的保守部分。在等温水浴中扩增 70 分钟后,含有 IBDV 的样品产生了预期的梯状产物,而其他病毒则没有产生产物。通过与逆转录聚合酶链反应(RT-PCR)和病毒分离进行比较,评估了 RT-LAMP 检测的敏感性和特异性。该检测方法比常规凝胶 RT-PCR 检测方法更为敏感。由于其特异性和简单性,RT-LAMP 检测方法可广泛应用于临床实验室,用于快速检测 IBVD。

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