Evaluation, standardization and development of molecular markers for the species-specific diagnosis of cutaneous leishmaniasis

• Grant number: 10/16963-4
• Support Opportunities: Regular Research Grants
• Start date: December 01, 2010
• End date: May 31, 2013
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal Investigator: Paulo Cesar Cotrim
• Grantee: Paulo Cesar Cotrim
• Host Institution: Instituto de Medicina Tropical de São Paulo (IMT). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: José Angelo Lauletta Lindoso

Abstract

We intend to analyze the sensitivity and specificity of five different primers for polymerase chain reaction (PCR) in order to consolidate a robust diagnostic method for identifying the species involved in Cutaneous Leishmaniasis (CL) in Brazil. We will compare the data collected from the results obtained with classical diagnosis methods of CL (direct examination, culture, serology, Montenegro skin-test), as well as clinical data of each patient. We intend to further validate this prospective test analyzing different reference species of parasite samples from other trypanosomatids, as well as samples from patients with related pathological processes. To this, four molecular targets were selected for the amplification: 1) conserved region of the minicircle kDNA, 2) region of rDNA internal transcribed spacers (ITS1rDNA), 3) region that encodes the enzyme glucose-6-phosphate dehydrogenase ( G6PD), and 4) region of the gene encoding the heat shock protein HSP70. It will also be assessed one-fifth and prospective target region containing a gene that encodes PRP-1, a membrane P-gluco protein. The PRP-1 was identified and described by our research group and is related to Pentamidine resistance in Leishmania. The development of this research project is a natural consequence of a multicenter project coordinated by FIOCRUZ (in final validation), where the methodologies for collecting clinical material, extraction and DNA amplification, and tests with restriction enzymes (RFLP ) for the identification of Leishmania species were standardized and validated by reference laboratories in Brazil, which had the participation of our research group (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

SATOW, MARCELA M.; YAMASHIRO-KANASHIRO, EDITE H.; ROCHA, MUSSYA C.; OYAFUSO, LUIZA K.; SOLER, RITA C.; COTRIM, PAULO C.; LINDOSO, JOSE ANGELO L.. APPLICABILITY OF kDNA-PCR FOR ROUTINE DIAGNOSIS OF AMERICAN TEGUMENTARY LEISHMANIASIS IN A TERTIARY REFERENCE HOSPITAL. Revista do Instituto de Medicina Tropical de São Paulo, v. 55, n. 6, p. 393-399, NOV-DEC 2013. (10/16963-4)