Abstract
BackgroundAmerican cutaneous leishmaniasis is an infectious dermatosis caused by protozoa of the genus Leishmania, which comprises a broad spectrum of clinical manifestations depending on the parasite species involved in the infections and the immunogenetic response of the host. The use of techniques for amplification of the parasites DNA based on polymerase chain reaction polymerase chain reaction and the recent application of combined techniques, such as high-resolution DNA dissociation, have been described as a viable alternative for the detection and identification of Leishmania spp. in biological samples. ObjectivesTo identify the Leishmania species using the polymerase chain reaction high-resolution DNA dissociation technique in skin biopsies of hospital-treated patients, and compare with results obtained by other molecular identification techniques. MethodsA retrospective study assessing patients with suspected American cutaneous leishmaniasis seen at a hospital in São Paulo/Brazil was conducted. The paraffin blocks of 22 patients were analyzed by polymerase chain reaction high-resolution DNA dissociation to confirm the diagnosis and identify the species. ResultsOf the 22 patients with suspected American cutaneous leishmaniasis, the parasite was identified in 14, comprising five cases (35.6%) of infection by L. amazonensis, four (28.5%) by L. braziliensis, two (14.4%) by L. amazonensis+L. infantum chagasi, two (14.4%) by L. guyanensis, and one (7.1%) by Leishmania infantum chagasi. In one of the samples, in which the presence of amastigotes was confirmed on histopathological examination, the polymerase chain reaction high-resolution DNA dissociation technique failed to detect the DNA of the parasite. Study limitationsThe retrospective nature of the study and small number of patients. ConclusionsThe method detected and identified Leishmania species in paraffin-embedded skin biopsies with a sensitivity of 96.4% and could be routinely used in the public health system.
Highlights
Leishmaniasis is a chronic infectious-parasitic disease caused by several species of protozoa of the genus Leishmania
Isolated cases of American cutaneous leishmaniasis (ACL) caused by Leishmania (L.) infantum chagasi have been reported in patients with or without HIV co-infection in the central-western and southwestern regions of the country.[4,5]
highresolution melting (HRM) analyses were performed at the end of each real-time polymerase chain reaction (PCR) reaction; the DNA dissociation profiles were generated by the High Resolution Melting software v. 3.0.1 (Life Technologies) and compared to profiles generated from DNA samples from reference strains of Leishmania spp. (L. (L.) infantum chagasi (MCER/BR/1981/M6445), L. (L.) major MHOM/IL/81/Friedlin), L. (L.)
Summary
Leishmaniasis is a chronic infectious-parasitic disease caused by several species of protozoa of the genus Leishmania. American cutaneous leishmaniasis is an infectious dermatosis caused by protozoa of the genus Leishmania, which comprises a broad spectrum of clinical manifestations depending on the parasite species involved in the infections and the immunogenetic response of the host. Objectives: To identify the Leishmania species using the polymerase chain reaction highresolution DNA dissociation technique in skin biopsies of hospital-treated patients, and compare with results obtained by other molecular identification techniques. Results: Of the 22 patients with suspected American cutaneous leishmaniasis, the parasite was identified in 14, comprising five cases (35.6%) of infection by L. amazonensis, four (28.5%) by L. braziliensis, two (14.4%) by L. amazonensis + L. infantum chagasi, two (14.4%) by L. guyanensis, and one (7.1%) by Leishmania infantum chagasi.
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