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首页> 外文期刊>Parasitology >Development and evaluation of different PCR-based typing methods for discrimination of Leishmania donovani isolates from Nepal.
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Development and evaluation of different PCR-based typing methods for discrimination of Leishmania donovani isolates from Nepal.

机译：开发和评估不同的基于PCR的区分尼泊尔利什曼原虫多形体分离株的分型方法。

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INTRODUCTION: Leishmania donovani, the causative agent of visceral leishmaniasis in the Indian subcontinent, has been reported to be genetically homogeneous. In order to support ongoing initiatives to eliminate the disease, highly discriminative tools are required for documenting the parasite population and dynamics. METHODS: Thirty-four clinical isolates of L. donovani from Nepal were analysed on the basis of size and restriction endonuclease polymorphisms of PCR amplicons from kinetoplast minicircle DNA, 5 nuclear microsatellites, and nuclear loci encoding glycoprotein 63, cysteine proteinase B, and hydrophilic acylated surface protein B. We present and validate a procedure allowing standardized analysis of kDNA fingerprint patterns. RESULTS: Our results show that parasites are best discriminated on the basis of kinetoplast minicircle DNA (14 genotypes) and 1 microsatellite defining 7 genotypes, while the remaining markers discriminated 2 groups or were monomorphic. Combination of all nuclear markers revealed 8 genotypes, while extension with kDNA data yielded 18 genotypes. CONCLUSION: We present tools that allow discrimination of closely related L. donovani strains circulating in the Terai region of Nepal. These can be used to study the micro-epidemiology of parasite populations, determine the geographical origin of infections, distinguish relapses from re-infection, and monitor the spread of particular variants.
机译：简介：据报道，印度次大陆内脏利什曼病的病原体利什曼原虫多诺万尼在遗传上是同质的。为了支持正在进行的消灭该疾病的计划，需要有高判别力的工具来记录寄生虫的数量和动态。方法：根据运动塑料微圆DNA，5个核微卫星和编码糖蛋白63，半胱氨酸蛋白酶B和亲水性酰化的核基因座的PCR扩增子的大小和限制性核酸内切酶多态性，分析了尼泊尔的L. donovani的34株临床分离株。表面蛋白B。我们提出并验证了一种程序，可以对kDNA指纹图谱进行标准化分析。结果：我们的结果表明，基于运动质体微圆DNA（14个基因型）和1个微卫星定义了7个基因型，对寄生虫的区分最佳，而其余标记区分2个组或单态性。所有核标记的组合揭示了8个基因型，而用kDNA数据扩展得到18个基因型。结论：我们提供了可以区分尼泊尔特莱地区流通的紧密相关的多诺氏乳杆菌菌株的工具。这些可用于研究寄生虫种群的微观流行病学，确定感染的地理起源，区分复发与再感染以及监测特定变异的传播。

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