The development and evaluation of a loop-mediated isothermal amplification (LAMP) method for detection of Babesia spp. infective to sheep and goats in China

文献类型： 外文期刊

第一作者： Guan, Guiquan

作者： Guan, Guiquan;Luo, Jianxun;Liu, Zhijie;Gao, Jinliang;Ma, Miling;Liu, Aihong;Dang, Zhisheng;Liu, Junlong;Ren, Qiaoyun;Jin, Yurong;Yin, Hong;Guan, Guiquan;Chauvin, Alain;Moreau, Emmanuelle;Inoue, Noboru;Thekisoe, Oriel M. M.;Sugimoto, Chihiro

作者机构：

关键词： Epidemiology: Population Studies;Parasitology;Methods and Techniques;Molecular Genetics: Biochemistry and Molecular Biophysics

期刊名称：EXPERIMENTAL PARASITOLOGY （ 影响因子：2.011； 五年影响因子：2.132 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： The loop-mediated isothermal amplification (LAMP) reaction is a method that amplifies with high sensitivity, efficiency, and rapidity, deoxyribonucleic acid (DNA) under isothermal condition in simple incubators. Two primer sets for the LAMP method were designed using the nucleotide sequences of 18S rRNA gene of Babesia sp. BQ1 (Lintan) and Babesia sp. Xinjiang-2005 isolated in China. The primers were used to detect parasite DNA extracted from infected blood and purified parasites by LAMP. The specific ladder bands were amplified from the autologous genomic DNA of two Babesia species, respectively, and did not cross-react with the genomic DNA of Theileria sp. China 1, Theileria sp. China 2, B. bovis, Theileria sp. (Japan) and sheep. The LAMP was sensitive enough to detect 0.02 pg and 0.2 pg genomic DNA of Babesia sp. BQ1 (Lintan) and Babesia sp. Xinjiang-2005, respectively, from 10-fold serially diluted samples corresponding to the amount of DNA present in 50 mu l of 0.000002% and 0.00002% parasitemic erythrocytes. Furthermore, DNA extracted from blood of intact (non-splenectomized) sheep experimentally infected with Babesia sp. BQ1 (Lintan) and Babesia sp. Xinjiang-2005 was amplified by the LAMP from week 1 to 9 and week 2 and 3 post-infection, respectively, demonstrating the high sensitivity of these primers. Of 365 samples collected from Gansu province, 14.3% (52/365) were positively detected by the LAMP. Of 145 samples collected on filter papers (Whatman) from the grazing sheep in Xinjiang province, 3.5% (5/145) were positive. These results show that the LAMP could be an alternative diagnostic tool for the detection of babesial infection in sheep and goats. (C) 2008 Elsevier Inc. All rights reserved.

分类号： Q18

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