Toxoplasma gondii infection: serological and molecular characterization in patients attended in public health service (SUS)

Abstract

Toxoplasmosis , a disease resulting from infection with Toxoplasma gondii , is clinically manifested in the brain forms , congenital and ocular . The prevalence of this disease varies in different areas around the world and depends on several factors . Epidemiological studies conducted in different Brazilian states in recent decades have revealed wide variability in rates of infection with T. gondii. In recent years our group has shown that rates of infection by T. gondii in pregnant women from the northwest region of São Paulo are approximately 65.0 % , the rate of congenital infection in São José do Rio Preto , SP is 2.3 % and ocular toxoplasmosis is approximately 27 % among patients suffering from eye diseases . The genetic characterization of T. gondii performed in human specimens reveals a great diversity and attempts to clarify the relationship between the different strains and human infection , including our region . The aim of this project is to investigate and characterize the infection by T. gondii in their serologic and molecular aspects in the northwest of São Paulo . Its specific objectives include: 1.Validar commercial serological tests used in the identification of anti -T . gondii in groups of patients with suspected or clinical diagnosis of ocular toxoplasmosis , congenital or gestational 2. To evaluate the applicability of these patients cPCR , Nested PCR and qPCR in biological samples ( peripheral blood and amniotic fluid ), 3. Genotypically characterized strains of T. gondii in the region using the PCR - RFLP method and establish relationship with the clinical features of patients affected ( Partnership abroad) . Will be selected serum samples and genomic DNA of patients with clinical diagnosis of ocular toxoplasmosis , pregnancy and congenital stored in the Laboratory of Immunogenetics , Department of Molecular Biology FAMERP whose epidemiological data of the same patients were collected . The methods commercial ELISA ( DiaSorin ) , ELFA ( Biomerieux ) and HALF (Abbott ) will be used for the identification of IgA , IgM and IgG anti- T. gondii and the identification of the genomic DNA of T. gondii is performed with the use of conventional PCR methods , nested PCR , Real Time PCR and PCR - RFLP . The results of serological and molecular analyzes will be compared . The statistical Ç2 ( with Yates correction ) , Fisher 's exact test and t test ( if applicable ) will be used in comparisons . Also be calculated odds ratio and confidence interval of 95 % , when necessary . The sensitivity, specificity and positive predictive values (PPV ) and negative (NPV ) will also be calculated for each employee molecular and serological testing in the analyzes. The results may contribute to understand the importance of adopting measures for preventing infection by T. gondii , and identify the type of strain that affect patients in the region . They can also guide programs clarification, education, prevention , epidemiology and even treatment of various forms of toxoplasmosis in the northwest of São Paulo .