RESUMO
We report a new endemic zone of cutaneous leishmaniasis (CL) in the central area of the State of Paraná (Municipality of Prudentópolis), in southern Brazil. This region was not previously considered endemic for CL, and this work constitutes the first report of CL endemicity there. Leishmaniasis was confirmed by smear, culture, and ELISA. Parasites were isolated and identified by random amplification of polymorphic DNA (PCR-RAPD). Phylogeographical analysis, based on two different criteria, was able to distinguish between RAPD profiles from different geographical regions. In total, 100 patients were diagnosed with leishmaniasis by culture and serology methods. The reported incidence rate was 4.32%. Of the 100 patients, 92% of the patients had single lesions, and 79.98% of these lesions were located on their limbs. The fact that 61% of patients were male rural workers points to an extradomiciliar type of transmission. In houses where human leishmaniasis was diagnosed, 29% of the dogs presented anti-Leishmania antibodies. A total of 1663 phlebotomines, representing 5 species, were captured in the studied area with CDC-like light minitraps. Lutzomyia intermedia s.l. was the most prevalent species (94.40%). The isolated parasites were grouped with Leishmania (V.) braziliensis. The epidemiological implications are discussed in the present article.
Assuntos
Doenças Endêmicas , Leishmania/isolamento & purificação , Leishmaniose Cutânea/epidemiologia , Adolescente , Adulto , Animais , Brasil/epidemiologia , Criança , Pré-Escolar , Análise por Conglomerados , Impressões Digitais de DNA , DNA de Protozoário/genética , Cães , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Geografia , Humanos , Incidência , Lactente , Leishmania/citologia , Leishmania/genética , Leishmania/imunologia , Masculino , Microscopia/métodos , Pessoa de Meia-Idade , Psychodidae/parasitologia , Técnica de Amplificação ao Acaso de DNA Polimórfico/métodos , População Rural , Adulto JovemRESUMO
The objective of the present study was to establish a polymerase chain reaction (PCR) technique for the diagnosis of cutaneous and mucocutaneous leishmaniosis from autochthonous cases in the state of Paraná in southern Brazil as well as imported cases. We sought to determine its utility and accuracy compared with smears and present culture methods. To standardize PCR samples, skin and mucosal punch biopsies from human lesions were performed on patients living in different regions of the Paraná state (76 cases) and other endemic areas of Brazil and Argentina (7 cases). For PCR standardization, two pairs of primers (MP1L/MP3H and B1/B2) were utilized for amplification of the conserved sequences in the minicircle of kinetoplast DNA (kDNA) for the Leishmania braziliensis complex. Two other primer pairs (b1/b2 and a1/a2) were species-specific for L. (V.) braziliensis and L. (V.) amazonensis, respectively. After differential diagnosis, eight patients had clinical diagnosis of the cutaneous ulcer changed to others pathologies such as syphilis, baso-cellular carcinoma, varicose ulcer, ecthyma and paracoccidioidomycosis. Of the 75 patients with cutaneous (CL) and mucocutaneous (MCL) lesions who provided samples, 47 (46 CL + 1 MCL) were diagnosed with leishmaniosis by smear and 57 (52 LC + 5 MCL) were diagnosed by culture methods. In contrast, our PCR technique presented higher accuracy when compared to the direct examination and culture of parasites. PCR is applicable both for CL where all 61 lesions were diagnosed, and MCL where 12 of 14 lesions were diagnosed. This molecular biology technique is also a faster and more specific diagnostic method compared with present parasitological procedures.