Centro di Referenza Nazionale per le Leishmaniosi, Palermo, Italy.
Transbound Emerg Dis. 2010 Apr;57(1-2):37-41. doi: 10.1111/j.1865-1682.2010.01131.x.
Different approaches are being developed to improve the differentiation of Leishmania genus using biochemical and molecular methods. In this study, 11 independent polymorphic microsatellites were used for the typing of strains of L. infantum isolated in Sicily. Polymerase chain reaction was employed to amplify the microsatellites contained in 12 DNA regions selected from among more investigated loci. A total of 51 isolates of L. infantum from dogs were tested by using the same locus panel. The products were successively analysed using an automatic sequence detector (ABI PRISM 3130 AB), to discover relevant microsatellite polymorphisms. It was possible to discriminate between MON-1 and non-MON-1 groups. Moreover, the method permitted to distinguish various genotypes of L. infantum isolates within each zymodema. Model- and distance-based analyses of the data set showed comparable results. The frequency of heterozygosity in the alleles analysed varied extremely between the different groups of isolates. As the method exhibits a high level of discrimination, it is suitable for characterization of closely related strains in epidemiological studies.
目前，人们正在开发不同的方法，通过生化和分子方法提高利什曼原虫属的分化能力。本研究使用 11 个独立的多态性微卫星对西西里岛分离的婴儿利什曼原虫菌株进行分型。聚合酶链反应用于扩增从更多研究的基因座中选择的 12 个 DNA 区域中包含的微卫星。使用相同的基因座面板对 51 株来自狗的婴儿利什曼原虫进行了测试。成功地使用自动序列检测仪（ABI PRISM 3130 AB）分析了产物，以发现相关的微卫星多态性。可以区分 MON-1 和非 MON-1 组。此外，该方法还可以区分每个酶谱内婴儿利什曼原虫分离株的不同基因型。基于模型和距离的数据分析结果相似。在不同的分离株组之间，分析的等位基因的杂合度频率差异极大。由于该方法具有较高的分辨能力，因此适合于在流行病学研究中对密切相关的菌株进行特征描述。
