Development and Evaluation of a Loop-Mediated Isothermal Amplification Method for Rapid Detection of Anaplasma ovis

文献类型： 外文期刊

第一作者： Luo, Jianxun

作者： Luo, Jianxun

作者机构：

期刊名称：JOURNAL OF CLINICAL MICROBIOLOGY （ 影响因子：5.948； 五年影响因子：5.345 ）

ISSN： 0095-1137

年卷期： 2011 年 49 卷 6 期

页码：

收录情况： SCI

摘要： Anaplasma ovis is an intraerythrocytic rickettsial pathogen of small ruminants. Loop-mediated isothermal amplification (LAMP) is a nucleic acid detection method in which the target DNA can be efficiently amplified with high specificity and sensitivity under isothermal conditions. In this study, a LAMP method was developed for the specific detection of A. ovis, using LAMP primers designed on the basis of the major surface protein 4 gene. LAMP was performed at 65 degrees C for 30 min. Its specificity was confirmed by successful amplification of several A. ovis isolates and through EcoRI restriction analysis of LAMP products. No cross-reactivity with the A. marginale Lushi isolate, Mycoplasma mycoides subsp. capri, Chlamydophila psittaci, Theileria ovis, T. luwenshuni, T. uilenbergi, or the Babesia sp. Xinjiang isolate was observed. Detection using the LAMP method was compared with that using conventional PCR in 227 field samples; LAMP demonstrated a sensitivity of 95.45%. In summary, LAMP is a specific, sensitive, and rapid test for the diagnosis of A. ovis infection, with the potential to be standardized as a detection method for A. ovis in areas of endemicity.

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