Molecular Identification of Parasites Causing Cutaneous Leishmaniasis in Panama

Abstract

Isolates from 475 cutaneous leishmaniasis (CL) patients from three endemic regions were studied by three typing techniques. The molecular analysis from lesion scrapings based on hsp70 PCR-restriction fragment length polymorphism (RFLP) showed that 78.1% (371/475) restriction patterns corresponded to Leishmania (Viannia) panamensis, 19% (90/475) to Leishmania (Viannia) guyanensis, and 3.0% (14/475) to Leishmania (Viannia) braziliensis. Promastigotes isolated by culture from lesions of 228 patients (48.0%, 228/475) were identified by multi-locus enzyme electrophoresis. Of them, 95.2% (217/228) were typified as L. (V.) panamensis, 1.3% (3/228) as L. (V.) guyanensis, 2.2% (5/228) as L. (V.) braziliensis, and 1.3% (3/228) as hybrids (L. [V.] braziliensis/L. [V.] panamensis). However, a partial sequencing analysis of the hsp70 gene from 77 selected samples showed 16.9% (13/77) typified as L. (V.) panamensis, 68.8% (53/77) as Leishmania (V.) sp., 1, 3.9% (3/77) as L. (V.) guyanensis, 1.3% (1/77) as L. (V.) braziliensis outlier, 2.6% (2/77) as Leishmania (Viannia) naiffi, 2.6% as (2/77) Leishmania (V.) sp., and 2 and 3.9% (3/77) hybrid isolates of L. (V.) braziliensis/L. (V.) guyanensis. These results confirm L. (V.) panamensis as the predominant species and cause of CL lesions in Panama and that L. (V.) guyanensis, L. (V.) braziliensis, and L. (V.) naiffi are circulating to a lower degree. Furthermore, the determination of parasite isolates belonging to atypical clusters and hybrid isolates suggests the circulation of genetic variants with important implications for the epidemiology and clinical follow-up of CL in Panama. No evidence of the existence of parasites of the Leishmania (Leishmania) mexicana complex in Panamanian territory was found in this study.

Sequencing of hsp70 for discernment of species from the Leishmania (Viannia) guyanensis complex from endemic areas in Colombia

“…MLEE analysis has questioned the distinction between L. (V.) panamensis and L. (V.) guyanensis since data did not indicate distinct monophyletic lines [43]. Furthermore, the zymodeme 2.3 strains sequenced in this work clustered with high support values together with strains sequenced in Panama by Miranda et al [34] and identi ed by the authors as Leishmania sp.1. Therefore, a close, if not identical, taxonomic identity is suggested.…”

“…Previous studies in Neotropical countries such as Ecuador, French Guiana, Brazil, Perú and Panama have reported the usefulness of hsp70 sequence analysis for Leishmania species identi cation [34,37,38,39,40].…”

“…To quantify diversity among co-occurring L. (V.) panamensis strains in each department, 31 sequences (25 from clinical isolates, ve from sand ies and one from a rodent) plus L. (V.) panamensis strains from Miranda et al [34] were analyzed to search for polymorphisms using DnaSP 6.10.03. Gap/missing bases were not considered, and invariable sites were removed.…”

“…Nine strains (codes 12581, 12615, 12578, 2183, 12563, 2198, 2168, 2173, and 2169) isolated from CL patients from the south paci c coast of Colombia typed as L.(V.) panamensis clustered with a group of reference sequences of Leishmania sp. from Panama [34]. Five out of those nine strains were also typed as L.(V.) panamensis zymodeme 2.3 using MLEE (Fig.…”

Characterization of Leishmania spp. Causing Cutaneous Lesions with a Negative Parasitological Diagnosis in Panama

“…Leishmania (V.) panamensis has been the main etiologic agent of CL reported in Panamá since it was accepted as a new species in 1972 [ 16 ]. However, recent studies have confirmed that at least one genetic variant of L. (V.) panamensis is also frequently found in Panamanian patients with CL [ 17 , 18 ]. These studies have also confirmed the presence of other species circulating in the country, such as L. (V.) braziliensis , L. (V.) guyanensis , and L. (V.) naiffi at low frequencies.…”