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Serological and molecular investigation of Toxoplasma gondii in blood donors

Abstract

Infection with Toxoplasma gondii, the etiological agent of toxoplasmosis, worldwide distribution and affects humans of all ages and ethnicities. The spread of the infection by different ways, including transfusion. In immunocompetent individuals, this parasite does not offer great risks, but in individuals imuncomprometidos involvement can be severe and result in high mortality. Objectives. The goal of this project is to detect the use of serological methods and describe the use of molecular methods to infection by T. gondii in blood donors. Specific objectives include: 1. Perform serological screening for anti-T. gondii (IgM and IgG) in suitable candidates for blood donation and identify their profiles [(IgM non-reactive, non-reactive IgG); (IgM, IgG non-reactive); (IgM, IgG reagent); (IgM nonreactive; IgG reagent)]. 2. Detecting the presence of genomic DNA of T. gondii in the peripheral blood of donors with the use of gene amplification methods (conventional PCR and real-time PCR). 3. Characterize the donors with positive PCR, the genotypes of T. gondii with the use of molecular markers. 4. Track units of blood that are experiencing positive for IgM and CRP (cn PCR and RT-PCR) and their receptors with the use of these serological and molecular methods. Patients and methods. Will be selected at least 2000 blood donors are able to donate at the Blood Center of São José do Rio Preto, São Paulo State. For each donor will be filled with a plug and epidemiological data are collected two samples of peripheral blood at the time of donation. One will be used for the detection of antibodies (IgM and IgG) anti-T. gondii, using the ELISA method. The other will be used for genomic DNA extraction using commercial kits. The gene amplification of T. gondii will be performed by conventional PCR (cnPCR) and real-time PCR (qPCR). The positive samples are genotyped by PCR using the markers SAG1, SAG2 (50-and 30-SAG2, alt.SAG2) SAG3, BTUB, GRA6, C22-8, c29-2, L358, and apical PK1. As the serological profile, obtained from ELISA, blood donors will be separated into four groups (G1, G2, G3 and G4). G1 will be included in the donor profile serological IgM and IgG non-reactive. In G2, are included those with IgM and IgG reagent; G3 in those IgM and IgG non-reactive, and G4 nonreactive IgM and IgG reagent. All these groups will be compared with the result of PCR for T. gondii. The results obtained and the PCR will be compared by using the Ç2 test and Fisher's exact test, when appropriate. Also be calculated Odds Ratio values and confidence intervals at 95%. Expected results. It is hoped that the results to be achieved with this project will contribute to meeting the T. gondii circulating in the peripheral blood of donors and to estimate the risk that the presence of such parasites provides patients requiring transfusion. (AU)

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Scientific publications (7)
(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)
DE MATTOS, LUIZ CARLOS; FERREIRA, ANA IARA COSTA; DE OLIVEIRA, KARINA YOUNAN; NAKASHIMA, FABIANA; BRANDAO, CINARA CASSIA. The Potential Contribution of ABO, Lewis and Secretor Histo-Blood Group Carbohydrates in Infection by Toxoplasma gondii. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 11, JUN 18 2021. Web of Science Citations: 0.
MIOLA, MARCOS PAULO; COLOMBO, TATIANA ELIAS; FACHINI, ROBERTA MARIA; RICCI-JUNIOR, OCTAVIO; BRANDAO DE MATTOS, CINARA CASSIA; DE MATTOS, LUIZ CARLOS. Anti-A and anti-A,B monoclonal antisera with high titers favor the detection of A weak phenotypes. TRANSFUSION AND APHERESIS SCIENCE, v. 59, n. 5 OCT 2020. Web of Science Citations: 0.
MURATA, FERNANDO HENRIQUE ANTUNES; PREVIATO, MARIANA; FREDERICO, FABIO BATISTA; BARBOSA, AMANDA PIRES; NAKASHIMA, FABIANA; DE FARIA, JR., GERALDO MAGELA; SILVEIRA CARVALHO, APARECIDA PERPETUO; MEIRA STREJEVITCH, CRISTINA DA SILVA; PEREIRA-CHIOCCOLA, VERA LUCIA; CASTIGLIONI, LILIAN; DE MATTOS, LUIZ CARLOS; SIQUEIRA, RUBENS CAMARGO; BRANDAO DE MATTOS, CINARA CASSIA. Evaluation of Serological and Molecular Tests Used for the Identification of Toxoplasma gondii Infection in Patients Treated in an Ophthalmology Clinic of a Public Health Service in Sao Paulo State, Brazil. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 9, FEB 7 2020. Web of Science Citations: 4.
NAKASHIMA, FABIANA; PARDO, VALQUIRIA SOUSA; MIOLA, MARCOS PAULO; ANTUNES MURATA, FERNANDO HENRIQUE; PADUAN, NATALIA; LONGO, STEFANI MIQUELINE; BRANDAO DE MATTOS, CINARA CASSIA; PEREIRA-CHIOCCOLA, VERA LUCIA; RICCI, OCTAVIO; DE MATTOS, LUIZ CARLOS. Serum IgG Anti-Toxoplasma gondii Antibody Concentrations Do Not Correlate Nested PCR Results in Blood Donors. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 9, JAN 14 2020. Web of Science Citations: 0.
ANTUNES MURATA, FERNANDO HENRIQUE; FERREIRA, MARINA NEVES; PEREIRA-CHIOCCOLA, VERA LUCIA; JUNQUEIRA FRANCO SPEGIORIN, LIGIA COSENTINO; MEIRA-STREJEVITCH, CRISTINA DA SILVA; GAVA, RICARDO; SILVEIRA-CARVALHO, APARECIDA PERPETUO; DE MATTOS, LUIZ CARLOS; BRANDAO DE MATTOS, CINARA CASSIA; GRP, FAMERP TOXOPLASMA RES. Evaluation of serological and molecular tests used to identify Toxoplasma gondii infection in pregnant women attended in a public health service in Sao Paulo state, Brazil. DIAGNOSTIC MICROBIOLOGY AND INFECTIOUS DISEASE, v. 89, n. 1, p. 13-19, SEP 2017. Web of Science Citations: 2.
ANTUNES MURATA, FERNANDO HENRIQUE; FERREIRA, MARINA NEVES; CAMARGO, NATALIA SAHYOUN; SANTOS, GABRIELA SORIA; JUNQUEIRA FRANCO SPEGIORIN, LIGIA COSENTINO; SILVEIRA-CARVALHO, APARECIDA PERPETUO; PEREIRA-CHIOCCOLA, VERA LUCIA; DE MATTOS, LUIZ CARLOS; BRANDAO DE MATTOS, CINARA CASSIA. Frequency of anti-Toxoplasma gondii IgA, IgM, and IgG antibodies in high-risk pregnancies, in Brazil. Revista da Sociedade Brasileira de Medicina Tropical, v. 49, n. 4, p. 512-514, JUL-AUG 2016. Web of Science Citations: 3.
FERNANDO HENRIQUE ANTUNES MURATA; MARINA NEVES FERREIRA; NATÁLIA SAHYOUN CAMARGO; GABRIELA SORIA SANTOS; LÍGIA COSENTINO JUNQUEIRA FRANCO SPEGIORIN; APARECIDA PERPÉTUO SILVEIRA-CARVALHO; VERA LUCIA PEREIRA-CHIOCCOLA; LUIZ CARLOS DE MATTOS; CINARA CÁSSIA BRANDÃO DE MATTOS. Frequency of anti- Toxoplasma gondii IgA, IgM, and IgG antibodies in high-risk pregnancies, in Brazil. Revista da Sociedade Brasileira de Medicina Tropical, v. 49, n. 4, p. 512-514, 2016-08-00.

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