Anticorpos anti-trypanosoma cruzi como preditivos de infecção por leishmania infantum

Leishmania infantum and Trypanosoma cruzi are trypanosomatids of medical importance and are, respectively, the etiologic agents of visceral leishmaniasis (VL) and Chagas disease (CD) in Brazil. People infected with L. infantum or T. cruzi may develop asymptomatically, enabling the transmission of pathogens through blood transfusion and / or organs. The assessment of the infection by T. cruzi is included among the tests performed for screening blood donors in Brazil, however, there is no availability of tests for Leishmania. Serological tests for T. cruzi are very sensitive, but not specific, and may have cross-reactions with other microorganisms. Thus, the aim of this study was to determine the prevalence of Leishmania infection in blood donors and assess whether the serological test for T. cruzi detect L. infantum. Among the 300 blood samples from donors, discarded in 2011, 61 were T. cruzi positive, 203 were from donors with other infections and 36 were from handbags with low blood volume, but without infection. We also assessed 144 samples from donors without infections and able to donate blood, totaling 444 subjects. DNA was extracted from blood samples of all to perform quantitative PCR (qPCR) to detect Leishmania DNA. The buffy coat obtained from all samples was grown in Schneider medium supplemented and NNN. All samples were evaluated for the presence of anti-Leishmania antibody. The serological results indicate a percentage of 22% of Leishmania infection in blood samples obtained from discarded bags. A total of 60% of samples positive in ELISA for T. cruzi were negative by IFI, used as confirmatory test, ie 60% false positive for Chagas. Among these samples false positive for Chagas, 72% were positive by ELISA for Leishmania characterizing the occurrence of cross reaction between serologic assays. Of the 300 cultures performed, 18 grew parasites that were typed by qPCR and specific isoenzymes, found the species Leishmania infantum crops. Among the 18 cultures, 4 were purged from scholarships for low volume and all negative serology blood bank, thus demonstrating that there is a real risk of Leishmania transmission via transfusion. It is concluded that in an area endemic for leishmaniasis in Brazil, serological diagnosis performed to detect infection by T. cruzi among blood donors can identify infection by L. infantum and although cause false positive for Chagas, this cross-reactivity reduces the risk of Leishmania infection via blood transfusion, since tests are not applied specific detection of the parasite. In this way, there remains the need to discuss the implementation of a specific serological screening test for Leishmania in endemic countries such as Brazil

Anticorpos anti-trypanosoma cruzi como preditivos de infecção por leishmania infantum

Leishmania infantum and Trypanosoma cruzi are trypanosomatids of medical importance and are, respectively, the etiologic agents of visceral leishmaniasis (VL) and Chagas disease (CD) in Brazil. People infected with L. infantum or T. cruzi may develop asymptomatically, enabling the transmission of pathogens through blood transfusion and / or organs. The assessment of the infection by T. cruzi is included among the tests performed for screening blood donors in Brazil, however, there is no availability of tests for Leishmania. Serological tests for T. cruzi are very sensitive, but not specific, and may have cross-reactions with other microorganisms. Thus, the aim of this study was to determine the prevalence of Leishmania infection in blood donors and assess whether the serological test for T. cruzi detect L. infantum. Among the 300 blood samples from donors, discarded in 2011, 61 were T. cruzi positive, 203 were from donors with other infections and 36 were from handbags with low blood volume, but without infection. We also assessed 144 samples from donors without infections and able to donate blood, totaling 444 subjects. DNA was extracted from blood samples of all to perform quantitative PCR (qPCR) to detect Leishmania DNA. The buffy coat obtained from all samples was grown in Schneider medium supplemented and NNN. All samples were evaluated for the presence of anti-Leishmania antibody. The serological results indicate a percentage of 22% of Leishmania infection in blood samples obtained from discarded bags. A total of 60% of samples positive in ELISA for T. cruzi were negative by IFI, used as confirmatory test, ie 60% false positive for Chagas. Among these samples false positive for Chagas, 72% were positive by ELISA for Leishmania characterizing the occurrence of cross reaction between serologic assays. Of the 300 cultures performed, 18 grew parasites that were typed by qPCR and specific isoenzymes, found the species Leishmania infantum crops. Among the 18 cultures, 4 were purged from scholarships for low volume and all negative serology blood bank, thus demonstrating that there is a real risk of Leishmania transmission via transfusion. It is concluded that in an area endemic for leishmaniasis in Brazil, serological diagnosis performed to detect infection by T. cruzi among blood donors can identify infection by L. infantum and although cause false positive for Chagas, this cross-reactivity reduces the risk of Leishmania infection via blood transfusion, since tests are not applied specific detection of the parasite. In this way, there remains the need to discuss the implementation of a specific serological screening test for Leishmania in endemic countries such as Brazil

International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients.

A century after its discovery, Chagas disease still represents a major neglected tropical threat. Accurate diagnostics tools as well as surrogate markers of parasitological response to treatment are research priorities in the field. The purpose of this study was to evaluate the performance of PCR methods in detection of Trypanosoma cruzi DNA by an external quality evaluation.

Leishmania spp. and/or Trypanosoma cruzi diagnosis in dogs from endemic and nonendemic areas for canine visceral leishmaniasis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Screening for anemia in Red Cross blood donor clinics

Accurate screening for anemia at Red Cross blood donor clinics is essential to maintain a safe national blood supply. Despite the importance of identifying anemia correctly by measurement of hemoglobin or hematocrit (hemoglobin/hematocrit) there is no consensus regarding the efficacy of the current two stage screening method which uses the Readacrit$\sp{\rm tm}$ microhematocrit in conjunction with copper sulfate.^ A cross-sectional study was implemented in which hemoglobin/hematocrit was measured, with the present method and four new devices, on 504 prospective blood donors at a Canadian Red Cross permanent blood donor clinic in London, Canada. Concurrently gathered, venous and capillary blood samples were tested by each device and compared to Coulter S IV$\sp{\rm tm}$ determined venous standard readings. Instrument hemoglobin/hematocrit means were statistically calibrated to the standard ones in order to appraise systematic deviations from the standard. Classification analysis was employed to assess concordance between each instrument and the standard when classifying prospective donors as anemic or non-anemic. This was done both when each instrument was used alone (single stage) and when copper sulfate was used as a preliminary screen (two stage) and simulated over a range of anemia prevalences. The Hemoximeter$\sp{\rm tm}$ and Compur M1000$\sp{\rm tm}$ devices had the highest correlations of hemoglobin measurements with the standard ones for both capillary (n.s.) and venous blood (p $<$.05). Analysis of variance (anova) also showed them to be the most accurate (p $<$.05), as did both single and two stage classification analysis, therefore, they are both recommended. There was a smaller difference between instruments for two stage than for single stage screening; therefore instrument choice is less crucial for the former. The present method was adequate for two stage screening as tested but simulations showed that it would discriminate poorly in populations with a higher prevalence of anemia. The Stat-crit and Readacrit, which measure hematocrit, became less accurate at crucial low hematocrit levels. In light of this finding and the introduction of new, effective and easy to use hemoglobin measuring instruments, the continued use of hematocrit as a surrogate for hemoglobin, is not recommended. ^

Synthesis and in vitro activity of limonene derivatives against Leishmania and Trypanosoma

The synthesis and in vitro activity of R(+)-Limonene derivatives against Leishmania and Trypanosoma cruzi strains are reported. Seven compounds have shown better in vitro activity against Leishmania (V.)braziliensis than the standard drug pentamidine. Additionally, we have identified two promising new anti-T. cruzi limonene derivatives. (C) 2010 Elsevier Masson SAS. All rights reserved.

Estudo Farmacognóstico e Screening Biológico de Solanum lycocarpum St. Hill (Solanaceae)

Pós-graduação em Ciências Farmacêuticas - FCFAR

Serological cross-reactivity of Trypanosoma cruzi, Ehrlichia canis, Toxoplasma gondii, Neospora caninum and Babesia canis to Leishmania infantum chagasi tests in dogs

Introduction: The aim of this study was to evaluate the serological cross-reactivity between Leishmania sp. and other canine pathogens. Methods: Positive serum samples for Ehrlichia canis, Babesia canis, Toxoplasma gondii, Neospora caninum and Trypanosoma cruzi were tested using three serological methods enzyme linked immunosorbent assay (ELISA), indirect immunofluorescent antibody test (IFAT) and Kalazar Detect™, for canine visceral leishmaniasis. Results: Of the 57 dog samples tested, 24 (42.1%) tested positive using one of the three serological methods: 10/57 (17.5%) for ELISA, 11/57 (19.3%) for IFAT and 3/57 (5.3%) for Kalazar Detect™. Conclusions: Our results demonstrated that the presence of other infectious agents may lead to cross-reactivity on leishmaniasis serological tests.

Participação de gambás e cães domiciliados como reservatórios de Leishmania infantum e Trypanosoma cruzi georreferenciados nos municípios da Regional de Saúde de Três Lagoas - MS

Pós-graduação em Doenças Tropicais - FMB

Avaliação da reatividade cruzada entre antígenos de Leishmania spp e Trypanosoma cruzi na resposta sorológica de cães pela técnica de imunofluorescência indireta (RIFI)

Visceral Leishmaniasis (VL) is a zoonosis caused by Leishmania spp. protozoa. Dog is the main parasite's reservoir, especially in urban areas. Indirect Immunofluorescent Antibody Test (IFAT) is commonly used for canine visceral leishmaniasis (CVL) diagnosis, by detection of antibodies against the parasite. However, Trypanosoma cruzi (T.cruzi) cross reactions can occur in serological diagnosis. In order to evaluate the occurrence of cross reactions between antigens of Leishmania spp. and T.cruzi, 150 blood samples of CVL epidemiological inquiry positive dogs were collected and sera were tested by IFAT. Dogs were culled at Zoonosis Control Center of Bauru (SP), an endemic area for VL. Another 150 dogs' blood samples were collected in Botucatu (SP), a non endemic area for LV, and sera were also tested by IFAT. Serum samples of dogs from Bauru were positive, both for Leishmania spp. and for T. cruzi, showing high cross-reactions incidence, reinforcing the need of diagnosis confirmation by other tests. Serum samples of dogs from Botucatu demonstrated low prevalence of positive results by IFAT, both for Leishmania spp. and T.cruzi. However, despite the low number of positive serological results for these parasites, the epidemiological and serological investigation for LV and Chagas disease in dogs from Botucatu must be kept, considering the importance of dog as domestic reservoir for both parasites.

Avaliação diagnóstica para Leishmania spp. e Trypanosoma cruzi em gatos domésticos procedentes da associação protetora dos animais do município de Ilha Solteira, SP, Brasil

Pós-graduação em Doenças Tropicais - FMB

Infecção por Leishmania spp. e/ou por Trypanosoma cruzi em cães provenientes de área endêmica e não endêmica para leishmaniose canina

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Profile of Trypanosoma cruzi Reactivity in a Population at High Risk for Endemic Pemphigus Foliaceus (Fogo Selvagem)

Fogo Selvagem (FS) is an autoimmune bullous disease with pathogenic IgG autoantibodies recognizing desmoglein 1 (Dsg1), a desmosomal glycoprotein. In certain settlements of Brazil, a high prevalence of FS (3%) is reported, suggesting environmental factors as triggers of the autoimmune response. Healthy individuals from endemic areas recognize nonpathogenic epitopes of Dsg1, and exposure to hematophagous insects is a risk factor for FS. Fogo selvagem and Chagas disease share some geographic sites, and anti-Dsg1 has been detected in Chagas patients. Indeterminate Chagas disease was identified in a Brazilian Amerindian population of high risk for FS. In counterpart, none of the FS patients living in the same geographic region showed reactivity against Trypanosoma cruzi. The profile of anti-Dsg1 antibodies showed positive results in 15 of 40 FS sera and in 33 of 150 sera from healthy individuals from endemic FS sites, and no cross-reactivity between Chagas disease and FS was observed.

kDNA gene signatures of Trypanosoma cruzi in blood and oesophageal mucosa from chronic chagasic patients

Trypanosoma cruzi presents a high degree of intraspecific variability, with possible implications for the pathogenesis of Chagas disease. The aim of this study was to evaluate T cruzi kDNA minicircle gene signatures using the low-stringency single-specific-primer PCR technique in both peripheral blood and oesophageal. mucosa from chronic chagasic patients, with or without megaesophagus, atone or in combination with cardiopathy and megacolon. It was not possible to identify a uniform pattern of shared bands between blood and oesophageal mucosa samples from individuals with the same clinical. form or mixed forms, suggesting multiple T. cruzi infections with differential tissue tropism. Thus, the results indicate that there is an intense intraspecific variability in the hypervariable regions of T cruzi kDNA, which has so far made it impossible to correlate the genetic profile of this structure with the clinical manifestations of Chagas disease. (C) 2008 Royal. Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. AIL rights reserved.