Abstract Seven isolates from patients with American cutaneous leishmaniasis in the Amazon region of Brazil were phenotypically suggestive of Leishmania (Viannia) guyanensis/L. (V.) shawi hybrids. In this work, two molecular targets were employed to check the hybrid identity of the putative hybrids. Heat shock protein 70 (hsp70) gene sequences were analyzed by three different polymerase chain reaction (PCR) approaches, and two different patterns of inherited hsp70 alleles were found. Three isolates presented heterozygous L. (V.) guyanensis/L. (V.) shawi patterns, and four presented homozygous hsp70 patterns involving only L. (V.) shawi alleles. The amplicon sequences confirmed the RFLP patterns. The high-resolution melting method detected variant heterozygous and homozygous profiles. Single-nucleotide polymorphism genotyping/cleaved amplified polymorphic site analysis suggested a higher contribution from L. (V.) guyanensis in hsp70 heterozygous hybrids. Additionally, PCR-RFLP analysis targeting the enzyme mannose phosphate isomerase (mpi) gene indicated heterozygous and homozygous cleavage patterns for L. (V.) shawi and L. (V.) guyanensis, corroborating the hsp70 findings. In this communication, we present molecular findings based on partial informative regions of the coding sequences of hsp70 and mpi as markers confirming that some of the parasite strains from the Brazilian Amazon region are indeed hybrids between L. (V.) guyanensis and L. (V.) shawi.

中文翻译：

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分子工具证实天然利什曼原虫（Viannia）guyanensis / L。（V.）沙威杂种在巴西亚马逊地区引起皮肤利什曼病

摘要从巴西亚马逊地区的美国皮肤利什曼病患者中分离出的7株在表型上提示利什曼原虫（Viannia）guyanensis / L。（V.）沙威混合动力车。在这项工作中，两个分子靶标被用来检查推定杂种的杂种身份。通过三种不同的聚合酶链反应（PCR）方法分析了热休克蛋白70（hsp70）基因序列，发现了两种不同的遗传hsp70等位基因模式。三个分离株呈现杂合的L.（V.）Guyanensis / L。（V.）沙威模式，和四个呈现纯合的hsp70模式，仅涉及L.（V.）沙威等位基因。扩增子序列证实了RFLP模式。高分辨率解链方法检测到了杂合和纯合变异体。单核苷酸多态性基因分型/切割扩增的多态性位点分析表明，L.（V.）Guyanensis在hsp70杂合杂种中的贡献更大。此外，针对酶甘露糖磷酸异构酶（mpi）基因的PCR-RFLP分析表明，L。（V.）沙威和L.（V.）Guyanensis的杂合和纯合裂解模式，证实了hsp70的发现。在本交流中，我们基于hsp70和mpi编码序列的部分信息区域作为标记物，提供了分子发现，证实了来自巴西亚马逊地区的某些寄生虫菌株确实是L.（V.）guyanensis和L.（ V.）以甘露糖磷酸异构酶（mpi）基因为靶标的PCR-RFLP分析表明，L。（V.）shawi和L.（V.）Guyanensis的杂合和纯合切割方式，证实了hsp70的发现。在本交流中，我们基于hsp70和mpi编码序列的部分信息区域作为标记物，提供了分子发现，证实了来自巴西亚马逊地区的某些寄生虫菌株确实是L.（V.）guyanensis和L.（五）。以甘露糖磷酸异构酶（mpi）基因为靶标的PCR-RFLP分析表明，L。（V.）shawi和L.（V.）Guyanensis的杂合和纯合切割方式，证实了hsp70的发现。在本交流中，我们基于hsp70和mpi编码序列的部分信息区域作为标记物，提供了分子发现，证实了来自巴西亚马逊地区的某些寄生虫菌株确实是L.（V.）guyanensis和L.（ V.）