Abstract
Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the mannose phosphate isomerase (mpi) gene was applied to 134 skin samples collected from patients with cutaneous leishmaniasis (CL) in Peru for identification of the infecting parasite at the species level, and the results were compared with those of cytochrome b (cyt b) gene sequencing obtained in previous studies. Although most results (121/134) including 4 hybrids of Leishmania (Viannia) braziliensis and L. (V.) peruviana corresponded to those obtained in the previous study, PCR-RFLP analyses revealed the distribution of putative hybrid strains between L. (V.) peruviana and L. (V.) lainsoni in two samples, which has never been reported. Moreover, parasite strains showing discordance between kinetoplast and nuclear genes (kDNA and nDNA), so-called mito-nuclear discordance, were identified in 11 samples. Of these, six strains had the kDNAs of L. (V.) braziliensis or L. (V.) peruviana and nDNAs of L. (V.) guyanensis, and three strains had the kDNAs of L. (V.) shawi and nDNAs of L. (V.) braziliensis. The rest were identified as mito-nuclear discordance strains having kDNAs of L. (V.) braziliensis or L. (V.) peruviana and nDNAs of L. (V.) lainsoni, and kDNAs of L. (V.) lainsoni and nDNAs of L. (V.) braziliensis. The results demonstrate that Leishmania strains in Peru are genetically more complex than previously considered.
Highlights
Parasitic protozoa belonging to the genus Leishmania show marked epidemiologic and clinical diversity, causing a wide-range of human and animal diseases extending from localized, selflimiting cutaneous lesions, and severe diffuse and destructive mucocutaneous lesions, to disseminating visceral infection that is fatal in the absence of treatment
The results demonstrate that genetically complex Leishmania strains are present in Peru, highlighting the need to combine both nuclear DNA (nDNA) and Kinetoplast DNA (kDNA) targets to improve the validity of species identifications for full details of gene property variants
The application of PCR-RFLP to 134 skin samples collected from patients with cutaneous leishmaniasis (CL) in Peru (Fig 2) identified 5 Leishmania species [L. (V.) braziliensis, L. (V.) peruviana, L. (V.) guyanensis, L. (V.) lainsoni, and L (L.) amazonensis] and two putative hybrids [L. (V.) braziliensis/L. (V.) peruviana and L. (V.) peruviana/L. (V.) lainsoni] (Table 1)
Summary
Parasitic protozoa belonging to the genus Leishmania show marked epidemiologic and clinical diversity, causing a wide-range of human and animal diseases extending from localized, selflimiting cutaneous lesions, and severe diffuse and destructive mucocutaneous lesions, to disseminating visceral infection that is fatal in the absence of treatment. Combining nuclear DNA (nDNA) and kDNA markers has improved the power of molecular data to detect unexpected genetically complex strains with characteristics of hybrid and mito-nuclear discordance widely distributed in Ecuador [17] These recent findings have shown the necessity of updating the available data in its neighbouring country of Peru, with a similar eco-epidemiological situation, and searching for genetic recombination in Peruvian strains that were identified at the species level by kinetoplast cytochrome b (cyt b) gene sequence analysis in a previous study. The latter led to the identification of 7 species and 1 hybrid: Leishmania (Viannia) braziliensis, L. The latter led to the identification of 7 species and 1 hybrid: Leishmania (Viannia) braziliensis, L. (V.) peruviana, L. (V.) guyanensis, L. (V.) lainsoni, L. (V.) shawi, Leishmania (Leishmania) mexicana, L. (L.) amazonensis, and a hybrid of L. (V.) braziliensis/L. (V.) peruviana [18]
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