Loop-mediated isothermal amplification (LAMP) method based on two species-specific primer sets for the rapid identification of Chinese Babesia bovis and B. bigemina
Loop-mediated isothermal amplification (LAMP) method based on two species-specific primer sets for the rapid identification of Chinese Babesia bovis and B. bigemina
复制标题
基于两种物种特异性引物组的环介导等温扩增（LAMP）方法用于快速鉴定中国牛巴贝虫和二联巴贝虫。
DOI：
10.1016/j.parint.2012.07.004
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发表时间：
2012-12-01
影响因子：
1.9
通讯作者：
Luo, Jianxun
中科院分区：
文献类型：
--
作者：
Liu, Aihong;Guan, Guiquan;Luo, Jianxun
牛巴贝斯虫病是一种主要由牛巴贝斯虫和/或双线巴贝斯虫引起的由硬蜱传播的血液原虫疾病，在世界范围内具有显著的发病率和死亡率。这种病在中国的大部分地区都很普遍。然而，很难快速区分牛乳杆菌和双链球菌。为了检测和区分这些物种，开发了一个针对内部转录间隔区(ITS)基因特定序列的环介导等温扩增(LAMP)平台。特异性试验表明，该抗体与牛巴贝斯虫、环状泰勒虫、中华泰勒虫、瑟氏泰勒虫、边缘无浆虫等未见交叉反应，与牛血白细胞无交叉反应。LAMP方法对牛黄杆菌和双链杆菌的检测灵敏度均为0.1pg DNA，优于经典的PCR方法。使用中国实验感染和自然感染牛的血液样本，对该方法的诊断价值进行了评估。这些发现表明，巴贝斯虫种特异性LAMP试验在检测和区分巴贝斯虫物种方面可能有潜在的临床应用，特别是在巴贝斯虫病流行的国家。皇冠版权所有(C)2012由爱思唯尔爱尔兰有限公司出版。保留所有权利。
Bovine babesiosis is a tick-transmitted hemoprotozoan disease that is mainly caused by Babesia bovis and/or Babesia bigemina and is characterized by significant morbidity and mortality worldwide. This disease is widespread in most parts of China. However, it is difficult to rapidly discriminate between the B. bovis and B. bigemina species. To detect and distinguish these species, a loop-mediated isothermal amplification (LAMP) platform that targets specific sequences of the internal transcribed spacer (ITS) genes was developed. Specificity testing revealed that there was no cross-reaction with the other tick-borne parasites B. ovate, B. major, unnamed bovine Babesia, Theileria annulata, Theileria sinensis, Theileria sergenti, and Anaplasma marginale, or with bovine white blood cells. The sensitivity of the LAMP method was 0.1 pg DNA for both B. bovis and B. bigemina, which was superior to that of the classical PCR methods. This assay was evaluated for its diagnostic utility using blood samples collected from experimentally and naturally infected cattle in China. These findings indicate that the Babesia species-specific LAMP assay may have potential clinical application in the detection and differentiation of Babesia species, particularly in countries in which babesiosis is endemic. Crown Copyright (C) 2012 Published by Elsevier Ireland Ltd. All rights reserved.