Rapid identification and differentiation of Theileria sergenti and Theileria sinensis using a loop-mediated isothermal amplification (LAMP) assay

文献类型: 外文期刊

第一作者: Liu, Aihong

作者: Liu, Aihong;Guan, Guiquan;Du, Pengfei;Gou, Huitian;Liu, Zhijie;Ma, Milin;Ren, Qiaoyun;Liu, Junlong;Yang, Jifei;Li, Youquan;Niu, Qinli;Bai, Qi;Yin, Hong;Luo, Jianxun;Zhang, Jun

作者机构:

关键词: Theileria sergenti;T. sinensis;Identification;Differentiation;LAMP assays

期刊名称:VETERINARY PARASITOLOGY ( 影响因子:2.738; 五年影响因子:2.951 )

ISSN: 0304-4017

年卷期: 2013 年 191 卷 1-2 期

页码:

收录情况: SCI

摘要: The present study developed and validated a species-specific loop-mediated isothermal amplification (LAMP) assay for the rapid detection and discrimination of two benign bovine Theileria species - T. sergenti and T. sinensis. The LAMP assay is inexpensive and easy to perform and involves a rapid reaction-the amplification can be performed in 55 min or 50 min under isothermal conditions of 61 degrees C or 63 degrees C, respectively, by employing a set of four species-specific primer mixtures. The results can be checked using agarose gels. The optimal assay conditions, under which the assay exhibited with no cross-reaction with other closely related tick-borne parasites (T. annulata, Babesia bovis, B. bigemina, B. major, B. ovata, B. U. sp., Anaplasma marginale) or between the two Theileria species of interest, was established. The assay is approximately 10-fold more sensitive than the conventional specific PCR assay. The LAMP assay was validated using DNA from 6 standard stocks in the laboratory and was evaluated for its diagnostic utility using blood samples collected from experimentally and naturally infection cattle or yaks in China. These findings indicate that this Theileria species-specific LAMP assay may have potential clinical applications for the detection and differentiation of two benign bovine Theileria species - T. sergenti and T. sinensis, especially in endemic countries. (C) 2012 Elsevier B.V. All rights reserved.

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