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Determination of polymorphisms in genes that encode MBL proteins in patients with LV from Lisbon region and molecular identification of Leishmania parasite

Grant number: 14/09783-0
Support type:Scholarships abroad - Research Internship - Post-doctor
Effective date (Start): September 01, 2014
Effective date (End): November 30, 2014
Field of knowledge:Biological Sciences - Parasitology - Protozoology of Parasites
Principal Investigator:Paulo Eduardo Martins Ribolla
Grantee:Rita de Cassia Viveiros da Silveira
Supervisor abroad: Lenea Maria da Graça Campino
Home Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil
Local de pesquisa : Universidade Nova de Lisboa, Portugal  
Associated to the scholarship:12/22312-1 - Padronization of molecular techniques for routine diagnosis of visceral leishmaniasis and determination of polymorphisms in genes that encode MBL and LPL proteins in patients with LV from the Public Health Laboratory of Bauru-SP, BP.PD

Abstract

Leishmaiasis refers to a spectrum of infection, that includes cutaneous, muco-cutaneous and visceral forms, and is caused by the protozoan of the genus Leishmania. This disease is considered a major public health problem. Leismaniasis is common in tropical regions and, according to World Health Organization (WHO), affects 12 million people, with 350 million people at risk situation. Some common serologic methods are used as diagnostic assays, as immune enzymatic assay (ELISA) and indirect immunofluorescence assay (IFI), however, none of them have high sensibility and specificity; moreover, these serologic tests can present false-negative results, and cross reactions with other trypanosomatid parasites. Despite the necessity of a rapid diagnosis to the treatment of visceral leishmaniasis (VL), some biomarkers can be employed to predict the risk of VL development, assisting the treatment. It is known that polymorphisms of genes that code for the protein mannose-binding lectin (MBL) are associated with pathogen infection and progression of various diseases, such as LV, and also, this genotypes can be used to predict the risk of diseases development. Because of that, sponsored by a Brazilian research agency (FAPESP - Fundação de Amparo à Pesquisa do Estado de São Paulo), we are realizing MBL genotyping in patients suspected of leishmaniasis, from the state hospital of Bauru, in the Bauru city, which is an endemic city which may contribute to prognosis. To verify if MBL gene can be used as a marker of risk in different populations, we aim to initiate a study with a sample of metropolitan area of Lisbon, Portugal, which will be provided by the leishmaniasis group of the Institute of Hygiene and Tropical Medicine of Lisbon, that have as Principal Investigator Doctor Lenea Campino. The results could give us an indicative if MBL gene is related or not with VL in different populations. Moreover, it will be analyzed the genetic variability of Leishmania sp. parasite in samples of people from Bauru and Lisbon, applying the molecular characterization methodology PCR-RFLP, as molecular target the parasite mitochondrial DNA (kDNA). kDNA has been widely used as molecular target, both as tool in general or specific diagnosis of leishmaniosis, and as a tool in genotyping of parasite in specie and subgenera levels, assuring high specificity and sensibility, by presenting multiple copies. With this approach it may be possible to correlate the clinical response of patients with the genetic variability of the parasite. The group of Dr. Lenea Campino has extensive experience with genotyping by kDNA - PCR -RFLP, and its application will be of great value to the results of this project. Moreover the contribution to my professional and academic development will be immeasurable. (AU)