Genotyping of Trypanosoma cruzi in samples of immigrant patients living in Europe: correlation between Trypanosoma cruzi molecular epidemiology and the different clinical forms of Chagas Disease in the European continent

• Grant number: 19/11943-0
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Effective date (Start): September 01, 2019
• Effective date (End): August 31, 2020
• Field of knowledge: Health Sciences - Medicine - Medical Clinics
• Principal Investigator: José Antonio Marin-Neto
• Grantee: Maykon Tavares de Oliveira
• Supervisor: Israel Molina Romero
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Research place: Vall d'Hebron Research Institute (VHIR), Spain
• Associated to the scholarship: 18/22093-4 - "Genotyping of Trypanosoma cruzi in samples from patients with Chronic Chagas' Heart Disease'', BP.PD

Abstract

Chagas disease (CDh) etiological agent is the protozoan hemoflagellate called Trypanosoma cruzi. The specie has a genetic structure that allows the subdivision into six distinct genetic groups, TcI to TcVI, those have presented differences regarding the geographical distribution, main biological properties and drug susceptibility. However, the association between genetic variability and clinical forms of CDh, to date is not yet established. It is well know the prevalence of the TcII and TcVI DTUs in several regions of Latin America associated with both the domestic and wild cycle of Chagas' disease. However, little is known about the circulation of the different T. cruzi genotypes in Chagas' Latin Americans immigrants living on the European continent. This disease has become a serious public health problem in Europe, especially due to vertical transmission and blood banks with contaminated samples, once the usual control measures are not applied in Europe. Thus, this project aims to verify the genotypes of the parasites present in 400 patients with CDh, treated at the Infectious Diseases outpatient clinic of the Vall d'Hebron University Hospital in Barcelona, Spain, and to evaluate the existence of potential targets in the parasite genome in order to correlate the results with the parasites clinical evolution To achieve our main goal, epimastigote forms belonging to the parasite six genotypes will be maintained in exponential growth and used as controls reference profiles. Peripheral blood (20ml) will be collected from each patient, where 5 ml of blood will be added to Guanidine / EDTA for subsequent DNA extraction and molecular characterization of the parasites; 5 ml of whole blood will be used for DNA extraction and Real time PCR for Chagas' diseas diagnosis; and 10 ml for hemoculture to isolate the parasites. Clinical evaluations will be performed in all chronic Chagasic patients. Genotyping will be performed using conventional multilocus PCR, using intergenic amplification of the Spliced Leader gene utilizing the UTCC and TCac (SL-IRac) primers, D7 variable domain of the 24S± subunit of ribosomal RNA, intergenic region of the gene of the Spliced Leader using the primers AM1 and 1B and region of the A10 nuclear fragment trough semi-nested PCR. In addition to genotyping, a genome screening of the patients parasites isolated by blood culture will be performed to identify potential target genes that may serve as chronic disease prognostic markers. The correlation of the results found in the Chagasic immigrant patients, in association with data from the patients treated at the HCFMRP-USP, will be extremely important for the understanding of the biological and clinical aspects of T. cruzi in chronic patients, as well as understanding the molecular epidemiology of T. cruzi in non-endemic countries.

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)

DE OLIVEIRA, MAYKON TAVARES; SULLEIRO, ELENA; DA SILVA, MARIA CLAUDIA; SILGADO, AROA; DE LANA, MARTA; DA SILVA, JOAO SANTANA; MOLINA, ISRAEL; ANTONIO MARIN-NETO, J.. Intra-Discrete Typing Unit TcV Genetic Variability of Trypanosoma cruzi in Chronic Chagas' Disease Bolivian Immigrant Patients in Barcelona, Spain. FRONTIERS IN CARDIOVASCULAR MEDICINE, v. 8, MAY 20 2021. (16/25403-9, 18/22093-4, 19/11943-0)

TAVARES DE OLIVEIRA, MAYKON; SULLEIRO, ELENA; SILGADO GIMENEZ, AROA; DE LANA, MARTA; ZINGALES, BIANCA; SANTANA DA SILVA, JOAO; ANTONIO MARIN-NETO, J.; MOLINA, ISRAEL. Quantification of parasite burden of Trypanosoma cruzi and identification of Discrete Typing Units (DTUs) in blood samples of Latin American immigrants residing in Barcelona, Spain. PLoS Neglected Tropical Diseases, v. 14, n. 6, JUN 2020. (16/25403-9, 19/11943-0)