Levels of anti-B13 antibodies over time in a cohort of chronic infected by Trypanosoma cruzi. Its relationship with specific treatment and clinical status.

The immunodominant B13 protein of Trypanosoma cruzi is found on the surface of trypomastigotes and exhibits cross-reactivity with the human cardiac myosin heavy chain; for which antibodies against this parasitic antigen may be involved in the development of disease pathology. In a cohort of chronically T. cruzi-infected adults, undergoing trypanocidal treatment, or not, we, therefore, decided to evaluate the levels of anti-B13 antibodies (ELISA-B13) and its eventual relationship with heart complaints. Two hundred twenty-eight serum samples from 76 chronically infected adults with an average follow-up of 24 years were analyzed. Thirty of them had received trypanocidal treatment. Among treated patients, anti-B13 Ab levels in successive samples showed a significant decrease in reactivity as the years after treatment increased (ANOVA test, p = 0.0049). At the end of the follow-up, 36.7% became non-reactive for ELISA B13. Untreated patients did not have significant variations in the level of anti-B13 antibodies during follow-up. None of the treated patients had electrocardiographic changes compatible with chronic chagasic cardiomyopathy, whereas 21.7% of those undergoing no treatment did show such kind of pathological electrocardiogram tracings. ELISA-B13 was reactive in all cases with heart involvement. Among untreated patients, there were no significant differences in anti-B13 antibodies when comparing individuals without proven pathology with those with chronic chagasic cardiomyopathy. Although treatment with trypanocidal drugs was followed by decreased anti-B13 antibody levels, such assessment was unhelpful in differentiating the evolution of chronic chagasic heart disease.

Geographic distribution of Trypanosoma cruzi genotypes detected in chronic infected people from Argentina. Association with climatic variables and clinical manifestations of Chagas disease.

Chronic Chagas disease affects large number of people in Latin America where it remains one of the biggest public health problems. Trypanosoma cruzi is genetically divided into seven discrete typing units (DTUs), TcI-TcVI and Tcbat, and exhibits differential distribution across vectors, host and transmission cycles. Clinical manifestations (cardiac, digestive and / or neurological) vary according to the geographical region; and the DTUs more frequently found in any of the chronic form of the disease, indeterminate or clinical, are TcI, TcII, TcV and TcVI. However, why they have a particular geographical distribution and how they affect the development of Chagas disease is still unknown. In this study, we assessed the geographic distribution of T. cruzi genotypes detected in chronic infected people from 57 localities of endemic regions of Argentina and analyzed their association with climatic variables. The prevalent DTUs detected in the whole population were TcV (47.4%) and TcVI (66.0%). TcI and TcII were identified in 5.2% each. All DTUs were detected in single and mixed infections (78.4% and 21.6%, respectively). TcV was found in infected people from localities with significantly higher average annual temperature, seasonal temperature and annual temperature range than those infected with TcVI. When we evaluated the association of DTUs with clinical manifestations of Chagas disease, the probability of finding TcVI in subjects with chronic Chagas cardiomyopathy (CCC) was higher than other DTUs, but without reaching statistical significance. Moreover, the probability of finding TcV in those who have not developed the disease after 20 years of infection was significantly higher than in CCC, either if it was present as unique DTU (reciprocal OR=4.95 95%CI: 1.42 to 17.27) (p=0.0117) or if it was also part of mixed infections (reciprocal OR=3.375; 95%CI: 1.227 to 9.276) (p=0.0264). There was no difference in the distribution of TcI between asymptomatic people and those with clinical manifestations, while TcII appeared more frequently in CCC cases, but without statiscal significance.

Chagas congénito en atención primaria de la salud en la Provincia de Santa Fe / Congenital Chagas disease in primary health care in Santa Fe province

Actualmente, la transmisión transplacentaria es la vía más frecuente de infección por Trypanosoma cruzi. El diagnóstico y tratamiento temprano de hijos infectados evita el riesgo de desarrollar miocardiopatía y las niñas dejan de ser potenciales fuentes de transmisión congénita. En este estudio se evaluó el seguimiento de hijos de mujeres infectadas por T. cruzi en Centros de Salud de la provincia de Santa Fe. Se estudiaron 19 madres y sus 51 hijos. 45% (23/51) de los hijos no habían sido estudiados previamente, y de éstos 21/23 resultaron negativos mientras que dos niñas de 3 y 7 años estaban infectadas. Los 28 niños restantes ya habían sido estudiados en los Centros de Salud, siendo positivas dos gemelas de 22 meses y una niña de 9 años; los otros 25/28 hijos no estaban infectados. Un 47% (9/19) de las madres tenían como único antecedente la serología materna positiva, y de las 4 mujeres que transmitieron la infección, tres pertenecían a este grupo. La edad promedio de diagnóstico fue: 20±6 años en las madres y 7,4±6,7 años en los hijos. Se requieren estrategias sanitarias que favorezcan el estudio para la infección por T. cruzi en mujeres antes del embarazo y el seguimiento de todos los hijos para no perder la oportunidad de tratamiento

Transplacental transmission is currently the most frequent route of infection by Trypanosoma cruzi. Early diagnosis and treatment of infected children avoids the risk of developing cardiomyopathy, and girls are no longer potential sources of congenital transmission. This study evaluated the follow-up of children of women infected with T. cruzi in Primary Care Centres of the province of Santa Fe. Nineteen mothers and their 51 children were studied. Among the 51 children, 23 had no previous diagnosis (45%). Of these, 21 were negative while 2 girls, ages 3 and 7, were infected. The remaining 28 children already had a diagnosis at the Health Centres, with 2 twins of 22 months and a 9-year-old girl who were positive; the other 25 children were not infected. Among the 19 mothers, 9 (47%) had the positive maternal serology as the only antecedent. Of the 4 women who transmitted the infection, 3 belonged to this group. The average age of diagnosis was: 20 ± 6 years in mothers and 7.4 ± 6.7 years in children. Health strategies are required to promote the detection of infected women before pregnancy and the monitoring of all children so as not to miss the opportunity for treatment

Simple methodology to directly genotype Trypanosoma cruzi discrete typing units in single and mixed infections from human blood samples.

Different DNA markers to genotype Trypanosoma cruzi are now available. However, due to the low quantity of parasites present in biological samples, DNA markers with high copy number like kinetoplast minicircles are needed. The aim of this study was to complete a DNA assay called minicircle lineage specific-PCR (MLS-PCR) previously developed to genotype the T. cruzi DTUs TcV and TcVI, in order to genotype DTUs TcI and TcII and to improve TcVI detection. We screened kinetoplast minicircle hypervariable sequences from cloned PCR products from reference strains belonging to the mentioned DTUs using specific kDNA probes. With the four highly specific sequences selected, we designed primers to be used in the MLS-PCR to directly genotype T. cruzi from biological samples. High specificity and sensitivity were obtained when we evaluated the new approach for TcI, TcII, TcV and TcVI genotyping in twenty two T. cruzi reference strains. Afterward, we compared it with hybridization tests using specific kDNA probes in 32 blood samples from chronic chagasic patients from North Eastern Argentina. With both tests we were able to genotype 94% of the samples and the concordance between them was very good (kappa=0.855). The most frequent T. cruzi DTUs detected were TcV and TcVI, followed by TcII and much lower TcI. A unique T. cruzi DTU was detected in 18 samples meantime more than one in the remaining; being TcV and TcVI the most frequent association. A high percentage of mixed detections were obtained with both assays and its impact was discussed.

Trypanocide treatment among adults with chronic Chagas disease living in Santa Fe city (Argentina), over a mean follow-up of 21 years: parasitological, serological and clinical evolution.

The efficacy of treatment with nifurtimox and/or benznidazole among adults with chronic Chagas disease with no previous electrocardiographic disturbances was evaluated over a mean follow-up of 21 years, by means of conventional serology, xenodiagnosis, clinical examination, electrocardiograms and chest X-ray. One hundred and eleven patients, between 17 and 46 years old, were studied: 54 underwent treatment (nifurtimox 27, benznidazole 27) and 57 remained untreated (control group). Xenodiagnosis was performed on 65% of them: 36/38 of the treated and 9/34 of the untreated patients had previous positive xenodiagnosis. Post-treatment, 133 xenodiagnoses were performed on 41 patients, all resulting negative. In the control group, 29 xenodiagnoses were performed on 14 patients; 2 resulted positive. Sera stored during the follow-up were simultaneously analyzed through conventional serology tests (IHA; DA-2ME; IIF). The serological evolution in the treated group was: a) 37% underwent negative seroconversion (nifurtimox 11, benznidazole 9); b) 27.8% decreased titers (nifurtimox 9, benznidazole 6), 9 showed inconclusive final serology (nifurtimox 7, benznidazole 2); c) 35.2% remained positive with constant titers (nifurtimox 7; benznidazole 12). The control group conserved the initial antibody levels during the follow-up. In the clinical evolution, 2/54 (3.7%) of the treated and 9/57 (15.8%) of the untreated patients showed electrocardiographic disturbances attributable to Chagas myocardiopathy, with a statistically relevant difference (p<0.05). Treatment caused deparasitation in at least 37% of the chronically infected adults and a protective effect on their clinical evolution.

Trypanocide treatment among adults with chronic Chagas disease living in Santa Fe city (Argentina), over a mean follow-up of 21 years: parasitological, serological and clinical evolution

The efficacy of treatment with nifurtimox and/or benznidazole among adults with chronic Chagas disease with no previous electrocardiographic disturbances was evaluated over a mean follow-up of 21 years, by means of conventional serology, xenodiagnosis, clinical examination, electrocardiograms and chest X-ray. One hundred and eleven patients, between 17 and 46 years old, were studied: 54 underwent treatment (nifurtimox 27, benznidazole 27) and 57 remained untreated (control group). Xenodiagnosis was performed on 65 percent of them: 36/38 of the treated and 9/34 of the untreated patients had previous positive xenodiagnosis. Post-treatment, 133 xenodiagnoses were performed on 41 patients, all resulting negative. In the control group, 29 xenodiagnoses were performed on 14 patients; 2 resulted positive. Sera stored during the follow-up were simultaneously analyzed through conventional serology tests (IHA; DA-2ME; IIF). The serological evolution in the treated group was: a) 37 percent underwent negative seroconversion (nifurtimox 11, benznidazole 9); b) 27.8 percent decreased titers (nifurtimox 9, benznidazole 6), 9 showed inconclusive final serology (nifurtimox 7, benznidazole 2); c) 35.2 percent remained positive with constant titers (nifurtimox 7; benznidazole 12). The control group conserved the initial antibody levels during the follow-up. In the clinical evolution, 2/54 (3.7 percent) of the treated and 9/57 (15.8 percent) of the untreated patients showed electrocardiographic disturbances attributable to Chagas myocardiopathy, with a statistically relevant difference (p<0.05). Treatment caused deparasitation in at least 37 percent of the chronically infected adults and a protective effect on their clinical evolution.

Avaliamos a eficácia do nifurtimox e/ou benznidazol, durante 21 anos em média, em adultos chagásicos crônicos sem alterações eletrocardiográficas iniciais, mediante sorologia convencional, xenodiagnóstico, exames clínicos, eletrocardiográficos e radiografia do tórax. Estudamos 111 pacientes (17 a 46 anos): 54 foram tratados (27 com nifurtimox e 27 com benznidazol) e 57 formaram o grupo controle. Foram submetidos ao xenodiagnóstico 65 por cento dos pacientes estudados: 36/38 tratados e 9/34 do grupo controle com xenodiagnóstico positivo prévio. Após tratamento, foram realizados 133 xenodiagnósticos em 41 pacientes, sendo todos negativos. Foram realizados 29 xenodiagnósticos em 14 pacientes do grupo controle, 2 foram positivos. A sorologia convencional foi realizada em soros estocados durante o seguimento. Evolução sorológica. Grupo tratado: a) 37 por cento negativaram (nifurtimox 11, benznidazol 9); b) 27,8 por cento diminuíram a titulação (nifurtimox 9, benznidazol 6), 9 deles apresentaram sorologia final discordante (nifurtimox 7, benznidazol 2; c) 35,2 por cento permaneceram positivos com titulação constante (nifurtimox 7, benznidazol 12). Grupo controle: conservou os níveis iniciais de anticorpos durante o seguimento. Evolução clínica: 2/54 (3,7 por cento) pacientes tratados e 9/57 não tratados apresentaram alterações eletrocardiográficas atribuíveis a miocardiopatia chagásica. Diferenças estatisticamente significantes (p<0,05). O tratamento produziu efeito de combate ao parasita em pelo menos 37 por cento dos infetados crônicos adultos e efeito protetor na evolução clínica.