Evaluation of PCR technique efficiency with specific identification of the etiologic agent for canine visceral leishmaniasis in the city of Embu das Artes

• Grant number: 11/04487-6
• Support type: Scholarships in Brazil - Master
• Effective date (Start): August 01, 2011
• Effective date (End): June 30, 2013
• Field of knowledge: Biological Sciences - Parasitology - Protozoology of Parasites
• Principal researcher: Paulo Cesar Cotrim
• Grantee: Thaynan Fernandes Cunha Martins
• Home Institution: Instituto de Medicina Tropical de São Paulo (IMT). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

The classical diagnosis of canine visceral leishmaniasis (CVL) basically consists in the isolation of parasite in vitro, its identification by microscopic analysis and/or serological tests. It is possible to infer that these methods are difficult to be implemented, time-consuming and present low sensitivity and/or specificity and are incapable to differentiate the parasite species involved. With the development of molecular methods such as Polymerase Chain Reaction (PCR), the diagnosis of CVL became more accurate and easy to perform. The aim of this project is to evaluate the sensitivity and specificity of PCR in the diagnosis of CVL, using different tissues of dogs seropositive for leishmaniasis in the city of Embu das Artes - SP, and analyze the ability of this method to detecting the disease-causing agents involved in this infection . For that, three molecular targets were selected for amplification: 1) conserved region of minicircle kDNA, 2) region that encodes the glucose-6-phosphate dehydrogenase enzyme (G6PD), and 3) region of the gene encoding the heat shock protein hsp70. Standardizing the collection of biological samples, we intended to compare the results obtained in the PCR reactions with samples from different tissues, with the data obtained in the parasitological and serological tests, as well with the clinical data observed. (AU)