Garcia A L, Kindt A, Quispe-Tintaya K W, Bermudez H, Llanos A, Arevalo J, Bañuls A L, De Doncker S, Le Ray D, Dujardin J C
Centro Universitario de Medicina Tropical, Casilla 3119, Cochabamba, Bolivia.
Infect Genet Evol. 2005 Mar;5(2):109-16. doi: 10.1016/j.meegid.2004.07.003.
Multi-locus enzyme electrophoresis is the current gold standard for the genetic characterisation of Leishmania. However, this method is time-consuming and, more importantly, cannot be directly applied to parasites present in host tissue. PCR-based methods represent an ideal alternative but, to date, a multi-locus analysis has not been applied to the same sample. This has now been achieved with a sample of 55 neotropical isolates (Leishmania (Viannia) braziliensis, L. (V.) peruviana, L. (V.) guyanensis, L. (V.) lainsoni and L. (L.) amazonensis), using five different genes as targets, four of which encoded major Leishmania antigens (gp63, Hsp70, H2B and Cpb). Our multi-locus approach strongly supports the current taxonomy and demonstrates a highly robust method of distinguishing different strains. Within L. (V.) braziliensis, we did not encounter so far specific genetic differences between parasites isolated from cutaneous and mucosal lesions. Interestingly, results provided by each of the different antigen-genes in the species considered, were different, suggesting different selective pressures. Our work emphasises the need for a multi-disciplinary approach to study the clinical pleomorphism of leishmaniasis.
多位点酶电泳是目前用于利什曼原虫基因特征分析的金标准。然而，这种方法耗时较长，更重要的是，不能直接应用于宿主组织中的寄生虫。基于聚合酶链反应（PCR）的方法是一种理想的替代方法，但迄今为止，多位点分析尚未应用于同一样本。现在，通过对55株新热带分离株（巴西利什曼原虫（维氏亚属）、秘鲁利什曼原虫（维氏亚属）、圭亚那利什曼原虫（维氏亚属）、兰氏利什曼原虫（维氏亚属）和亚马逊利什曼原虫（利什曼亚属））的样本进行研究，以五个不同的基因作为靶点，其中四个基因编码主要的利什曼原虫抗原（gp63、热休克蛋白70（Hsp70）、组蛋白H2B（H2B）和半胱氨酸蛋白酶B（Cpb）），实现了多位点分析。我们的多位点方法有力地支持了当前的分类学，并证明了一种区分不同菌株的高度稳健的方法。在巴西利什曼原虫（维氏亚属）中，到目前为止，我们尚未发现从皮肤和黏膜病变中分离出的寄生虫之间存在特定的基因差异。有趣的是，在所研究的物种中，每个不同抗原基因提供的结果都不同，这表明存在不同的选择压力。我们的工作强调了采用多学科方法研究利什曼病临床多态性的必要性。
