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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  hsp 70 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 .

Molecular tools confirm natural Leishmania (Viannia) guyanensis/L. (V.) shawi hybrids causing cutaneous leishmaniasis in the Amazon region of Brazil