State Key Laboratory for Molecular Biology of Special Economic Animals, Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, China.
J Virol Methods. 2013 Feb;187(2):401-5. doi: 10.1016/j.jviromet.2012.11.012. Epub 2012 Nov 23.
Loop-mediated isothermal amplification (LAMP) method was discovered in the last decade but only used for the first time in the diagnosis of mink enteritis virus (MEV) infection in this study. The amplification could be completed within 60 min, under isothermal condition at 65°C, by employing a set of four primers targeting the VP2 gene of MEV. The LAMP was more sensitive than the conventional PCR, with a detection limit of 10(-1) median tissue culture infective doses (TCID(50))/ml per reaction, compared with 10 TCID(50)/ml for PCR analysis. No cross reactivity was observed for other related viruses, including canine distemper virus (CDV) and Aleutian mink disease parvovirus (AMDV). Eighty four of 230 clinical samples were found to be positive for MEV, which is higher than that determined by using the conventional PCR method (68). The results indicate the LAMP can be potentially used to determine MEV as a simple, rapid procedure. This assay would be an available alternative to PCR analysis for the diagnosis of MEV infection in mink, particularly in less well-equipped laboratories and in rural settings where resources are limited.
环介导等温扩增（LAMP）方法是在上个十年发现的，但在本研究中是首次用于貂肠炎病毒（MEV）感染的诊断。该扩增可以在 65°C 的等温条件下在 60 分钟内完成，采用针对 MEV 的 VP2 基因的四组引物。与常规 PCR 相比，LAMP 更敏感，检测限为 10(-1)中位数组织培养感染剂量（TCID(50)）/反应毫升，而 PCR 分析的检测限为 10 TCID(50)/ml。对于其他相关病毒，包括犬瘟热病毒（CDV）和阿留申病细小病毒（AMDV），没有观察到交叉反应。在 230 份临床样本中，有 84 份被检测为 MEV 阳性，高于使用常规 PCR 方法检测到的阳性率（68）。结果表明，LAMP 可用于确定 MEV 作为一种简单、快速的方法。与 PCR 分析相比，该检测方法可作为貂 MEV 感染的一种替代方法，特别是在设备较差和资源有限的农村地区的实验室。
