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Leishmania (Viannia) braziliensis genotypes identified in lesions of patients with atypical or typical manifestations of tegumentary leishmaniasis: Evaluation by two molecular markers
Authors:C Baptista  AO Schubach  CA Leal  FS Oliveira  CMV Rosalino  RS Pacheco
Institution:a Centro de Referência em Leishmanioses, Instituto de Pesquisa Clínica Evandro Chagas, IPEC/Fiocruz, Av. Brasil, 4365, Rio de Janeiro, Rio de Janeiro CEP 21040-900, Brazil
b Curso de Pós-Graduação em Pesquisa Clínica em Doenças Infecciosas, Instituto de Pesquisa Clínica Evandro Chagas, IPEC/Fiocruz, Av. Brasil, 4365, Rio de Janeiro, Rio de Janeiro CEP 21040-900, Brazil
c Laboratório de Sistemática Bioquímica, Instituto Oswaldo Cruz, Fiocruz, Av. Brasil, 4365, Rio de Janeiro, Rio de Janeiro CEP 21040-900, Brazil
d Laboratório de Imunoparasitologia, Instituto Oswaldo Cruz, Fiocruz, Av. Brasil, 4365, Rio de Janeiro, Rio de Janeiro CEP 21040-900, Brazil
Abstract:Analyses of MLEE, RAPD and LSSP-PCR were used to compare the panel of american tegumentary leishmaniasis (ATL) isolates obtained from lesions of patients with rare clinical manifestations of the disease and typical lesions. All of the 34 samples analyzed by MLEE demonstrated similar electromorphic profiles with Leishmania (Viannia) braziliensis reference strain. Through the RAPD analysis, nine genetic profiles (genotypes) were identified. LSSP-PCR corroborates the initial screening and phenetic analysis has grouped the isolates into two major clusters comprising the nine different genotypes. Prevalent genotype defined as LbmtDNAgen1 was detected in the largest number of isolates. There was no association between genotypes and clinical symptoms. However, two different genotypes could be identified in the initial (LbmtDNAGen9) and reactivated lesion (LbmtDNAGen3) of the same patient. Our results support the idea of a less pronounced genotypic diversity among L. (V.) braziliensis circulating in the State of Rio de Janeiro and demonstrate the useful application of these molecular markers in genetics variability studies.
Keywords:Leishmaniasis  Leishmania (Viannia)braziliensis  MLEE  multilocus enzyme electrophoresis  RAPD  random amplified polymorphic DNA  LSSP-PCR  low stringency single specific primer-polymerase chain reaction  kDNA  kinetoplast DNA  genetic variability
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