Diagnóstico de leptospirose por PCR e caracterização de isolados de Leptospira spp. por seqüenciamento do 16S rDNA e análise de VNTR / Diagnosis of leptospirosis by PCR and characterization of Leptospira spp. isolates by 16S rDNA sequencing and VNTR analysis

AUTOR(ES)
DATA DE PUBLICAÇÃO

2005

RESUMO

Leptospirosis, caused by bacteria of the genus Leptospira, is a direct anthropo-zoonotic infection with a geographical distribution throughout the world. Definitive diagnosis of leptospirosis has traditionally depended upon the isolation of leptospiras from clinical specimens or the demonstration of seroconversion in paired acute and convalescent serum samples. There is an urgent need for the development of new diagnostic strategies for leptospirosis. Conventional PCR assays have been developed, but they have limitations that have restricted their widespread use. In order to overcome these limitations, a nested PCR assay was developed using as target part of the lipL32 gene, which is conserved among pathogenic serovars of Leptospira. This approach was much more sensitive than conventional PCR. Furthemore, there are also difficulties associated with serological identification of Leptospira strains and for this reason we carried out molecular characterization of 10 Leptospira spp. strains isolated from human and animal clinical samples. The 16S rDNA sequencing allowed confirmation of the species, whereas serovar identification was only possible with the VNTR analysis. The analysis based on VNTR polymorphism provides a rapid as well as highly discriminating assay to characterize L. interrogans isolates at serovar level.

ASSUNTO(S)

bacterologia biotecnologia leptospira isolation vntr nested-pcr

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