RESUMO
La leishmaniasis cutánea se caracteriza por la inducción de una respuesta inmune celular y humoral, siendo los niveles de anticuerpos IgG e IgE indicadores serológicos del diagnóstico, la progresión y resolución de la enfermedad. En el presente estudio se estandarizaron dos pruebas de ELISA indirecto para determinar los niveles séricos de anticuerpos IgG o IgE contra antígeno soluble de Leishmania braziliensis (ASL). Se trabajó a partir de la cepa de L. braziliensis (MHOM/BR/75/M2903). En el ELISA IgE se hizo un pretratamiento del suero con anticuerpos antiIgG humano y Polietilenglicol (PEG); la dilución óptima de la muestra para ambos ELISA fue 1/21, la dilución del conjugado fue 1/500 para el ELISA IgG y 1/100 para el IgE. Para el ELISA IgG estandarizado se encontró un índice de correlación de Kappa de 1 (p<0,05), frente al ELISA comercial rBiopharm®. Con una sensibilidad del 100%; 100% de especificidad, 100% valor predictivo positivo y 100% valor predictivo negativo. Sin embargo ambos test de ELISA presentaron reacciones cruzadas con anticuerpos anti-Trypanosoma. La validez de los resultados demuestran que los ELISAs estandarizados pueden ser utilizados como herramienta de diagnóstico y seguimiento al tratamiento leishmanicida.
Cutaneous leishmaniasis is characterized by the induction of a cellular and humoral immune responses, at levels of IgG and IgE indicators serological diagnosis, progression and resolution of disease. In this study two indirect ELISA tests were standardized to determine serum levels of IgG or IgE antibodies against soluble Leishmania braziliensis antigen (ASL). We worked from the strain of L. braziliensis (MHOM / BR /75 / M2903). In the IgE ELISA pretreatment serum was done with anti-human IgG and Polyethylene glycol (PEG) antibodies; the optimal dilution of the sample for both ELISA was 1/21, the conjugate dilution was for 1/500 and 1/100 IgG ELISA for IgE. For standardized ELISA IgG index Kappa correlation of 1 (p <0.05) compared to commercial ELISA r-Biopharm® found. With a sensitivity of 100%; 100% specificity, 100% positive predictive value and negative predictive value 100%. However both ELISA test showed cross-reactions anti-trypanosome antibodies. The results demonstrate that both ELISA can be used as diagnostic and monitoring tool. Being necessary to continue studies to validate these procedures and identify other possible cross-reactions.
Assuntos
Humanos , Trypanosoma , Leishmania braziliensis , Ensaio de Imunoadsorção Enzimática , Assistência ao Convalescente , Anticorpos , Doença , Diagnóstico , Indicadores e ReagentesRESUMO
El Lupus Eritematoso Sistémico (LES) es una enfermedad autoinmune sistémica que afecta a diferentes órganos especialmente al riñón. Un gran número de estudios genéticos en diferentes grupos poblacionales han reportado asociaciones entre el polimorfismo en el intrón 16 del gen que codifica la enzima convertidora de la angiotensina (ECA) con la susceptibilidad a LES y a sus manifestaciones clínicas en particular a la nefropatía lúpica. El presente estudio pretende asociar el polimorfismo genotípico y alélico inserción/deleción (I/D) del gen ECA en pacientes bolivianos con susceptibilidad a LES y a las manifestaciones clínicas que presenta el paciente. Se incluyeron 87 pacientes lúpicos que cumplían con al menos cuatro criterios clínicos de la Sociedad Americana de Reumatología y 85 controles sanos. Todos los participantes dieron su consentimiento informado para participar en el estudio. Se detectaron los polimorfismos alélicos y genotípicos ECA I/I, I/D y D/D mediante la Reacción en Cadena de la Polimerasa (PCR). La frecuencia genotípica I/I, I/D y D/D en los pacientes lúpicos era 47.1, 48.3 y 4.6%; en el grupo control era 68.2, 31.8 y 0% respectivamente. En lúpicos la frecuencia alélica era del 71.3 y 28,7%; en controles 84.1 y 15.9% para los alelos I y D respectivamente. Se evidenció que el alelo inserción y sus variantes genotípicas I/I e I/D está sistemáticamente asociado con riesgo al desarrollo de complicaciones clínicas dermatológicas, osteomusculares o hematológicas. En el caso de nefropatía lúpica el alelo inserción es un factor de protección. El polimorfismo estudiado del gen ECA no mostro asociación de riesgo o protección con manifestaciones cardiopulmonares ni neurológicas.
Systemic Lupus Erythematosus (SLE) a systemic autoimmune disease, affects different organs, being kidneys the most frequently affected. Many studies in different population groups have reported associations between the intron 16 of the gene encoding angiotensin converting enzyme (ACE) polymorphism and SLE susceptibility and its clinical manifestations, in particular lupus nephritis. Associate genotypic and allelic insertion/deletion (I/D) of the ACE gene polymorphism in SLE patient's susceptibility and the clinical manifestations were the aims of the present study. 87 lupus patients who met at least four clinical criteria of the American Society of Rheumatology and 85 healthy controls where included. All participants gave their informed consent to participate in the study. Allelic and genotypic ACE I/I, I/D and D/D polymorphisms were detected by polymerase chain reaction (PCR). In patients s with SLE, the genotypic frequency of I/I, I/D and D/D were 47.1, 48.3 and 4.6%; and 68.2, 31.8 and 0% in control group respectively. Allele frequency in SLE represent: Insertion 71.3%, Deletion 28.7% and Insertion 84.1%, Deletion 15.9% in control group. Allele and genotypic variants I/I and I/D of the insertion allele were associated systematically with risk to dermatological, musculoskeletal and hematologic complications. Regarding lupus nephritis allele insertion is a protective factor. ACE gene polymorphism here studied has not been associated to risk or protection with cardiopulmonary or neurological clinical manifestations.
Assuntos
Humanos , Doenças Autoimunes , Reação em Cadeia da Polimerase , Lúpus Eritematoso Sistêmico , Polimorfismo Genético , Nefrite Lúpica , Consentimento Livre e EsclarecidoRESUMO
The Amazon region has recently been considered as endemic in Latin America. In Bolivia, the vast Amazon region is undergoing considerable human migrations and substantial anthropization of the environment, potentially renewing the danger of establishing the transmission of Chagas disease. The cases of human oral contamination occurring in 2010 in the town of Guayaramerín provided reasons to intensify research. As a result, the goal of this study was to characterize the species of sylvatic triatomines circulating in the surroundings of Yucumo (Beni, Bolivia), a small Amazonian city at the foot of the Andes between the capital (La Paz) and Trinidad the largest city of Beni. The triatomine captures were performed with mice-baited adhesive traps mostly settled in palm trees in forest fragments and pastures. Species were identified by morphological observation, dissection of genitalia, and sequencing of three mitochondrial gene fragments and one nuclear fragment. Molecular analysis was based on (i) the identity score of the haplotypes with GenBank sequences through the BLAST algorithm and (ii) construction of phylogenetic trees. Thirty-four triatomines, all belonging to the Rhodnius genus, of which two were adult males, were captured in palm trees in forest fragments and pastures (overall infestation rate, 12.3%). The morphology of the phallic structures in the two males confirmed the R. stali species. For the other specimens, after molecular sequencing, only one specimen was identified with confidence as belonging to Rhodnius robustus, the others belonged to one of the species of the Rhodnius pictipes complex, probably Rhodnius stali. The two species, R. robustus and R. stali, had previously been reported in the Alto Beni region (edge of the Amazon region), but not yet in the Beni department situated in the Amazon region. Furthermore, the difficulties of molecular characterization of closely related species within the three complexes of the genus Rhodnius are highlighted and discussed.
