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Montalvo A.M.,Institute Medicina Tropical Pedro Kour | Fraga J.,Institute Medicina Tropical Pedro Kour | Monzote L.,Institute Medicina Tropical Pedro Kour | Montano I.,Institute Medicina Tropical Pedro Kour | And 4 more authors.
Parasitology | Year: 2010

Introduction. Species typing in leishmaniasis gains importance in diagnostics, epidemiology, and clinical studies. A restriction fragment length polymorphism (RFLP) assay of PCR amplicons from a partial heat-shock protein 70 gene (hsp70) had been described for the New World, allowing identification of some species. Methods. Based on an initial in silico analysis of 51 hsp70 sequences, most of which we recently determined in the frame of a phylogenetic study, species-specific restriction sites were identified. These were tested by PCR-RFLP on 139 strains from 14 species, thereby documenting both inter- and intra-species variability. Results. Our assay could identify Leishmania infantum, L. donovani, L. tropica, L. aethiopica, L. major, L. lainsoni, L. naiffi, L. braziliensis, L. peruviana, L. guyanensis, and L. panamensis by applying 2 subsequent digests. L. mexicana, L. amazonensis, and L. garnhami did not generate species-specific restriction fragment patterns. Conclusion. Currently no assay is available for global Leishmania species discrimination. We present a universal PCR-RFLP method allowing identification of most medically relevant Old and New World Leishmania species on the basis of a single PCR, obviating the need to perform separate PCRs. The technique is simple to perform and can be implemented in all settings where PCR is available. Copyright © 2010 Cambridge University Press.


PubMed | Institute Medicina Tropical Pedro Kour
Type: Journal Article | Journal: Revista cubana de medicina tropical | Year: 2013

For the first time in Cuba the rapid serologic technologies available worldwide were evaluated for the screening and confirmation of human leptospirosis. As its clinical recognition is difficult, the fast etiological diagnosis is of vital importance. Sensitivity and specificity values higher than 90 % were obtained in the confirmatory systems, and of 85 % in the screening systems. Of the severe patients studied by these technologies 50 % were positive. 203 cases corresponding to 4 epidemic outbreaks were confirmed, as well as 12 sick persons vaccinated with vaxSPIRAL, which demonstrates the vaccine effectiveness (78.1%). We developed and applied a latex system for a rapid screening of leptospirosis (LeptoCuba), with an excellent sensitivity, specificity, reproducibility, and stability. The application of new technologies for the screening and fast confirmation of the disease allowed to increase the positivity and quality of the diagnosis from 2000 to 2006, strengthening the microbiological surveillance in the country.

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