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布氏锥虫:利用移动遗传元件的PCR分析进行锥虫菌株分型(MGE-PCR)

Trypanosoma brucei: trypanosome strain typing using PCR analysis of mobile genetic elements (MGE-PCR).

作者信息

Tilley A, Welburn S C, Fèvre E M, Feil E J, Hide G

机构信息

Centre for Tropical Veterinary Medicine, Royal (Dick) School of Veterinary Science, University of Edinburgh, Easter Bush, Roslin, Midlothian, Scotland EH25 9RG, UK.

出版信息

Exp Parasitol. 2003 May-Jun;104(1-2):26-32. doi: 10.1016/s0014-4894(03)00114-0.

Abstract

We describe the development of a single-primer amplification system, which uses the trypanosomal mobile genetic element RIME as a molecular marker for the differentiation of Trypanosoma brucei stocks. Using a well-characterised set of T. brucei stocks from southeast Uganda, Kenya and Zambia, we have evaluated the application of this technique, termed MGE-PCR (mobile genetic element PCR) for the typing of trypanosome strains. The technique revealed considerable variation between stocks and was sufficiently specific to amplify trypanosomal DNA in the presence of host DNA. The results showed a clear distinction between human-infective and non-human-infective stocks. Comparative studies on these stocks using markers for the human serum resistance associated (SRA) gene, which identifies human-infective stocks, demonstrated complete agreement between MGE-PCR derived groups and human-infectivity status. Furthermore, MGE-PCR detects high levels of variability within the T. b. brucei and T. b. rhodesiense groups and is therefore a powerful discriminatory tool for tracking individual T. brucei genotypes and strains.

摘要

我们描述了一种单引物扩增系统的开发,该系统使用锥虫移动遗传元件RIME作为分子标记来区分布氏锥虫株系。利用来自乌干达东南部、肯尼亚和赞比亚的一组特征明确的布氏锥虫株系,我们评估了这项被称为MGE-PCR(移动遗传元件PCR)的技术在锥虫菌株分型中的应用。该技术揭示了不同株系之间存在相当大的差异,并且具有足够的特异性,能够在宿主DNA存在的情况下扩增锥虫DNA。结果显示,人类感染性株系和非人类感染性株系之间有明显区别。使用与人血清抗性相关(SRA)基因的标记对这些株系进行的比较研究表明,MGE-PCR得出的分组与人类感染性状态完全一致,而SRA基因可识别出人类感染性株系。此外,MGE-PCR检测到布氏锥虫布鲁斯亚种和布氏锥虫罗德西亚亚种组内存在高度变异性,因此是追踪单个布氏锥虫基因型和菌株的强大鉴别工具。

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