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.
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.
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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