Development of loop-mediated isothermal amplification (LAMP) assay for rapid diagnosis of ovine theileriosis in China

文献类型： 外文期刊

第一作者： Liu, Zhijie

作者： Liu, Zhijie;Salih, Diaeldin A.;Ahmed, Jabbar S.;Seitzer, Ulrike;Liu, Zhijie;Hou, Junlin;Luo, Jianxun;Yin, Hong;Bakheit, Mohammed A.

作者机构：

期刊名称：PARASITOLOGY RESEARCH （ 影响因子：2.289； 五年影响因子：2.403 ）

ISSN： 0932-0113

年卷期： 2008 年 103 卷 6 期

页码：

收录情况： SCI

摘要： Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid detection method in which the target deoxyribonucleic acid (DNA) can be efficiently amplified with high specificity and sensitivity under isothermal conditions using a set of either four or six specific primers. In this study, we have identified a conserved sequence for Theileria luwenshuni (UTRlu8) and for T. uilenbergi (UTRu6) suitable for designing a set of six primers for the simultaneous detection by LAMP of these pathogens causing theileriosis in sheep and goats in China. LAMP was performed at 63 degrees C, and the amplified DNA was detectable within 15 min. The specificity of the reaction was confirmed through EcoRI restriction enzyme digestion analysis and sequencing. The assay was proven sensitive since specific amplification was obtained from 0.1 pg DNA of T. luwenshuni or T. uilenbergi. The LAMP assay was evaluated by testing 86 field samples in comparison to the reverse line blot method, showing a sensitivity and specificity of 66.0% and 97.4%, respectively. These results indicate that the LAMP assay is rapid and simple to run, cost effective, sensitive, and specific and has potential usefulness for application in diagnostics of and epidemiological studies on T. luwenshuni and T. uilenbergi infection of small ruminants.

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