Loop-mediated isothermal amplification (LAMP) assays for detection of Theileria parva infections targeting the PIM and p150 genes
Loop-mediated isothermal amplification (LAMP) assays for detection of Theileria parva infections targeting the PIM and p150 genes
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DOI：
10.1016/j.ijpara.2009.07.004
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发表时间：
2010-01-01
影响因子：
4
通讯作者：
Inoue, Noboru
中科院分区：
文献类型：
--
作者：
Thekisoe, Oriel M. M.;Rambritch, Natasha E.;Inoue, Noboru
我们已经建立了两种环介导的等温扩增(LAMP)分析方法，用于检测微小泰勒氏菌，它是东海岸热(ECF)的病原体，东海岸热是非洲东部、中部和南部的一种重要的牛疾病。这些检测针对的是多态免疫优势分子(PIM)和P150 LAMP基因。针对每个基因靶标的引物集由6个引物组成，每组都识别目标基因上的8个不同区域，以提供对T parva的高度特异性检测。每套引物的检测下限为1fg，相当于PIM和P150T parva基因的一个拷贝。这些PIM和P150 LAMP引物组合扩增了来自肯尼亚、南非、坦桑尼亚、卢旺达、乌干达和布隆迪等不同国家的牛和水牛的细小毛滴虫分离株的DNA，表明它们有能力检测来自不同国家的细小毛滴虫。这些LAMP分析具有简单、快速和成本效益高的优点，是分子流行病学研究和资源贫乏的ECF流行国家监测控制计划的良好候选者。(C)2009澳大利亚寄生虫学会有限公司出版。保留所有权利。
We have developed two loop-mediated isothermal amplification (LAMP) assays for the detection of Theileria parva, the causative agent of East Coast fever (ECF), an economically important cattle disease in eastern, central and southern Africa. These assays target the polymorphic immunodominant molecule (PIM) and p150 LAMP genes. The primer set for each gene target consists of six primers, and each set recognises eight distinct regions on the target gene to give highly specific detection of T parva. The detection limit of each primer set is 1 fg, which is equivalent to one copy of the PIM and p150 T parva genes. These PIM and p150 LAMP primer sets amplify DNA of T parva isolates from cattle and buffalo from different countries including Kenya, South Africa, Tanzania, Rwanda, Uganda and Burundi, indicating their ability to detect T. parva from different countries. With the advantages of simplicity, rapidity and cost effectiveness, these LAMP assays are good candidates for molecular epidemiology studies and for monitoring control programs in ECF-endemic, resource poor countries. (C) 2009 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.