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Título: A PCR assay for the identification of Leishmania species of the Viannia subgenus
Palabras clave: Biología
Leishmania
leishmaniasis
diagnostic
restriction fragments length polymorphism
polymerase chain reaction
B-tubulin gene
Editorial: Sociedad Venezolana de Microbiología
Descripción: We have identified a novel DNA sequence of 500 bp (B-500-DNA) on the Leishmania (Viannia) subgenus, located in the intergenic region of one of the loci of the B-tubulin gene family. The sequence analysis showed that this sequence has no homology to any other sequence described so far, including the B-tubulin gene. We improved a specific B-500-PCR assay, which generated a PCR product of 375 bp for total genomic DNA from Leishmania strains belonging to the L. (Viannia) subgenus. In contrast, no amplification was found when using genomic DNA from species of L. (Leishmania) subgenus or other organisms. Under our PCR conditions, the lower detection limit was 1 fg when a purified DNA clone (pLgB4), which contains one copy of the B-500-DNA sequence, was used. The B-500-DNA PCR assay confirmed the preliminary diagnosis of cutaneous leishmaniasis in clinical samples in which the Montenegro skin test was positive and parasite cultures were negative. The analytical specificity and the sensitivity of the PCR assay provide a tool for epidemiological studies of the disease.
Other Identifiers: http://www.redalyc.org/articulo.oa?id=199421829012
Aparece en las Colecciones:Revista de la Sociedad Venezolana de Microbiología

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