The identification, diversity and prevalence of trypanosomes in field caught tsetse in Tanzania using ITS-1 primers and fluorescent fragment length barcoding
The identification, diversity and prevalence of trypanosomes in field caught tsetse in Tanzania using ITS-1 primers and fluorescent fragment length barcoding
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DOI：
10.1016/j.meegid.2007.07.013
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发表时间：
2008-07-01
影响因子：
3.2
通讯作者：
Gibson, W. C.
中科院分区：
文献类型：
--
作者：
Adams, E. R.;Hamilton, P. B.;Gibson, W. C.
我们报告的发展，两个通用的，基于PCR的方法，取代以前使用的多个物种特异性PCR检测，以确定锥虫物种进行个别采采蝇。第一种方法是基于核糖体RNA（rRNA）基因座ITS-1区域PCR产物的种间大小差异。在第二种方法中，18 S和28 S rRNA基因内的多个片段的长度变化通过使用荧光引物的PCR扩增进行测定;随后通过使用自动DNA测序仪准确且快速地测定产物的大小。这两种方法都被用来识别在坦桑尼亚的塔兰吉雷和塞伦盖蒂国家公园以及Msubugwe沿海森林保护区对锥虫感染的采采蝇进行大规模实地研究期间收集的样本。随着时间的推移和两个不同的领域季节锥虫患病率的波动进行了讨论。除了以更快的速度、精度和灵敏度促进锥虫物种的鉴定外，这些通用系统还使我们能够鉴定两种新的锥虫物种。(C)2007 Elsevier B. V.保留所有权利。
We report on the development of two generic, PCR-based methods, which replace the multiple species-specific PCR tests used previously to identify the trypanosome species carried by individual tsetse flies. The first method is based on interspecies size variation in the PCR product of the ITS-1 region of the ribosomal RNA (rRNA) locus. In the second approach, length variation of multiple fragments within the 18S and 28S rRNA genes is assayed by PCR amplification with fluorescent primers; products are subsequently sized accurately and rapidly by the use of an automated DNA sequencer. Both methods were used to identify samples collected during large-scale field studies of trypanosome-infected tsetse in Tanzania in the National Parks of Tarangire and Serengeti, and the coastal forest reserve of Msubugwe. The fluctuations of trypanosome prevalence over time and two different field seasons are discussed. As well as facilitating the identification of trypanosome species with increased speed, precision and sensitivity, these generic systems have enabled us to identify two new species of trypanosome. (C) 2007 Elsevier B.V. All rights reserved.