Trypanosome identification in wild tsetse populations in Tanzania using generic primers to amplify the ribosomal RNA ITS-1 region
Trypanosome identification in wild tsetse populations in Tanzania using generic primers to amplify the ribosomal RNA ITS-1 region
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DOI：
10.1016/j.actatropica.2006.10.002
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发表时间：
2006-11-01
期刊：
影响因子：
2.7
通讯作者：
Gibson, W. C.
中科院分区：
文献类型：
--
作者：
Adams, E. R.;Malele, I. I.;Gibson, W. C.
采采蝇在非洲传播多种锥虫，其中一些是对人类和牲畜造成重大医疗和社会经济影响的病原体。鉴定锥虫对于评估特定采采蝇种群造成的疾病风险至关重要。我们开发了一种单一的通用PCR检测，以取代以前用于鉴定单个采采蝇携带的锥虫病种的多种物种特异性PCR检测。在通用PCR测试中，核糖体RNA重复区内部转录间隔区（ITS-1） PCR产物的种间大小变化使物种识别成为可能。该方法用于鉴定坦桑尼亚两个地区野生捕获的蝇类中肠样本中的锥虫。通过对扩增的ITS-1区域进行测序和/或物种特异性PCR检测来验证鉴定。该方法可快速准确地对大量野外样品进行鉴定。虽然物种特异性测试不能识别以前未知的物种，但通用测试可以通过扩增产物的独特大小来识别新物种。因此，即使没有获得该新物种的任何分离物，我们也可以收集其分布，流行和与其他锥虫共发生的数据。分子和生态图谱的结合将有助于该物种的分离和全面生物学特性的研究。(c) 2006 Elsevier B.V.版权所有
Tsetse flies transmit many species of trypanosomes in Africa, some of which are human and livestock pathogens of major medical and socio-economic impact. Identification of trypanosomes is essential to assess the disease risk posed by particular tsetse populations. We have developed a single generic PCR test to replace the multiple species-specific PCR tests used previously to identify the trypanosorne species carried by individual tsetse flies. In the generic PCR test, inter-species size variation in the PCR product of the internal transcribed spacer (ITS-1) region of the ribosomal RNA repeat region enables species identification. The test was applied to identify trypanosomes in midgut samples stored on FTA cards from wild-caught flies in two regions of Tanzania. Identifications were verified by sequencing the amplified ITS-1 region and/or species-specific PCR tests. The method facilitated the identification of large numbers of field samples quickly and accurately. Whereas species- specific tests are incapable of recognising previously unknown species, the generic test enabled a new species to be identified by the unique size of the amplified product. Thus, even without access to any isolate of this new species, we could collect data on its distribution, prevalence and co-occurrence with other trypanosomes. The combined molecular and ecological profiles should facilitate the isolation and full biological characterization of this species in the future. (c) 2006 Elsevier B.V. All rights reserved.