RESUMO
Toxocarosis is a zoonotic disease caused by the ingestion of infective eggs of Toxocara spp. The diagnosis is based on the detection of antibodies in serum or other biological fluids. One of the current serological techniques for the diagnosis of toxocariasis is ELISA using excretory - secretory antigens of third stage larvae (ES/L3). These antigens are glycoproteins, which originate in the secretory organs of the parasite and are non species-specific. Sera from patients with other helminthiases and non- parasitic diseases were used to evaluate the specificity of ELISA using the excretory - secretory antigen (ES/L3). The reactivity of these sera was between 11 and 70%. Western blot using patients' sera revealed that the glycoprotein triplet having a molecular weight of 120 kDa was responsible for cross-reactivity. With these results, and for the purpose of purifying the antigen, ion exchange chromatography was performed. When the sera from patients with various parasitic and non-parasitic diseases were analyzed with the purified antigen ES/ L3, they were only reactive between 10 to 20%. The sensitivity of the ELISA test determined by program Epidat 3.0 for the two antigens was 100%, but the following differences in specificity were observed: 84% for the total antigen ES/L3 and 99% for purified ES/L3. Using the ES/L3 purified antigen, it can be considered that the reactive sera, with compatible symptoms correspond to patients who are or were parasitized with Toxocara canis.
Assuntos
Antígenos de Helmintos/sangue , Toxocaríase/sangue , Toxocaríase/diagnóstico , Humanos , Testes Sorológicos/métodosRESUMO
La toxocariosis es una zoonosis causada por la ingestión de huevos infectivos de Toxocara spp. El diagnóstico de la enfermedad se basa en la detección de anticuerpos en el suero u otros fluidos biológicos. La técnica serológica más utilizada es el ELISA, que usa como antígeno los productos de excreción-secreción de larvas de tercer estadio (ES/L3). Estos productos antigénicos son glicoproteínas que se originan en los órganos secretorios del parásito y no son específicos de especie. Para evaluar la especificidad de la técnica de ELISA con el antígeno ES/L3, se emplearon sueros de personas con otras helmintiasis y con patologías no parasitarias. Se observó que estos sueros presentaron reactividad entre el 11 y el 70 % de los casos. El Western blot con suero de los mismos pacientes reveló que la glicoproteína que corresponde al triplete de 120 kDa fue la más inespecífica. Teniendo en cuenta estos resultados y con el propósito de purificar el antígeno se realizó una cromatografía de intercambio iónico. Cuando se analizaron los sueros de los pacientes con diferentes enfermedades parasitarias y no parasitarias con el antígeno ES/L3 purificado, solo fueron reactivos entre un 10 y un 20 % de ellos. La sensibilidad del test de ELISA determinada por el programa Epidat 3. 0 para los dos antígenos fue del 100 %, pero se observaron diferencias en la especificidad: para el antígeno ES/L3 total esta fue del 84 % y para el ES/L3 purificado del 99 %. Empleando el antígeno ES/L3 purificado se puede considerar que los sueros que son reactivos, en presencia de una sintomatología compatible, corresponden a pacientes que fueron o están parasitados con Toxocara canis.
Toxocarosis is a zoonotic disease caused by the ingestion of infective eggs of Toxocara spp. The diagnosis is based on the detection of antibodies in serum or other biological fluids. One of the current serological techniques for the diagnosis of toxocariasis is ELISA using excretory - secretory antigens of third stage larvae (ES/L3). These antigens are glycoproteins, which originate in the secretory organs of the parasite and are non species-specific. Sera from patients with other helminthiases and non- parasitic diseases were used to evaluate the specificity of ELISA using the excretory - secretory antigen (ES/L3). The reactivity of these sera was between 11 and 70%. Western blot using patients' sera revealed that the glycoprotein triplet having a molecular weight of 120 kDa was responsible for cross-reactivity. With these results, and for the purpose of purifying the antigen, ion exchange chromatography was performed. When the sera from patients with various parasitic and non-parasitic diseases were analyzed with the purified antigen ES/ L3, they were only reactive between 10 to 20%. The sensitivity of the ELISA test determined by program Epidat 3. 0 for the two antigens was 100%, but the following differences in specificity were observed: 84% for the total antigen ES/L3 and 99% for purified ES/L3. Using the ES/L3 purified antigen, it can be considered that the reactive sera, with compatible symptoms correspond to patients who are or were parasitized with Toxocara canis.
Assuntos
Humanos , Antígenos de Helmintos/sangue , Toxocaríase/sangue , Toxocaríase/diagnóstico , Testes Sorológicos/métodosRESUMO
La toxocariosis es una zoonosis causada por la ingestión de huevos infectivos de Toxocara spp. El diagnóstico de la enfermedad se basa en la detección de anticuerpos en el suero u otros fluidos biológicos. La técnica serológica más utilizada es el ELISA, que usa como antígeno los productos de excreción-secreción de larvas de tercer estadio (ES/L3). Estos productos antigénicos son glicoproteínas que se originan en los órganos secretorios del parásito y no son específicos de especie. Para evaluar la especificidad de la técnica de ELISA con el antígeno ES/L3, se emplearon sueros de personas con otras helmintiasis y con patologías no parasitarias. Se observó que estos sueros presentaron reactividad entre el 11 y el 70 % de los casos. El Western blot con suero de los mismos pacientes reveló que la glicoproteína que corresponde al triplete de 120 kDa fue la más inespecífica. Teniendo en cuenta estos resultados y con el propósito de purificar el antígeno se realizó una cromatografía de intercambio iónico. Cuando se analizaron los sueros de los pacientes con diferentes enfermedades parasitarias y no parasitarias con el antígeno ES/L3 purificado, solo fueron reactivos entre un 10 y un 20 % de ellos. La sensibilidad del test de ELISA determinada por el programa Epidat 3. 0 para los dos antígenos fue del 100 %, pero se observaron diferencias en la especificidad: para el antígeno ES/L3 total esta fue del 84 % y para el ES/L3 purificado del 99 %. Empleando el antígeno ES/L3 purificado se puede considerar que los sueros que son reactivos, en presencia de una sintomatología compatible, corresponden a pacientes que fueron o están parasitados con Toxocara canis.(AU)
Toxocarosis is a zoonotic disease caused by the ingestion of infective eggs of Toxocara spp. The diagnosis is based on the detection of antibodies in serum or other biological fluids. One of the current serological techniques for the diagnosis of toxocariasis is ELISA using excretory - secretory antigens of third stage larvae (ES/L3). These antigens are glycoproteins, which originate in the secretory organs of the parasite and are non species-specific. Sera from patients with other helminthiases and non- parasitic diseases were used to evaluate the specificity of ELISA using the excretory - secretory antigen (ES/L3). The reactivity of these sera was between 11 and 70%. Western blot using patients sera revealed that the glycoprotein triplet having a molecular weight of 120 kDa was responsible for cross-reactivity. With these results, and for the purpose of purifying the antigen, ion exchange chromatography was performed. When the sera from patients with various parasitic and non-parasitic diseases were analyzed with the purified antigen ES/ L3, they were only reactive between 10 to 20%. The sensitivity of the ELISA test determined by program Epidat 3. 0 for the two antigens was 100%, but the following differences in specificity were observed: 84% for the total antigen ES/L3 and 99% for purified ES/L3. Using the ES/L3 purified antigen, it can be considered that the reactive sera, with compatible symptoms correspond to patients who are or were parasitized with Toxocara canis.(AU)
Assuntos
Humanos , Antígenos de Helmintos/sangue , Toxocaríase/sangue , Toxocaríase/diagnóstico , Testes Sorológicos/métodosRESUMO
Toxocarosis is a zoonotic disease caused by the ingestion of infective eggs of Toxocara spp. The diagnosis is based on the detection of antibodies in serum or other biological fluids. One of the current serological techniques for the diagnosis of toxocariasis is ELISA using excretory - secretory antigens of third stage larvae (ES/L3). These antigens are glycoproteins, which originate in the secretory organs of the parasite and are non species-specific. Sera from patients with other helminthiases and non- parasitic diseases were used to evaluate the specificity of ELISA using the excretory - secretory antigen (ES/L3). The reactivity of these sera was between 11 and 70
. Western blot using patients sera revealed that the glycoprotein triplet having a molecular weight of 120 kDa was responsible for cross-reactivity. With these results, and for the purpose of purifying the antigen, ion exchange chromatography was performed. When the sera from patients with various parasitic and non-parasitic diseases were analyzed with the purified antigen ES/ L3, they were only reactive between 10 to 20
. The sensitivity of the ELISA test determined by program Epidat 3.0 for the two antigens was 100
, but the following differences in specificity were observed: 84
for the total antigen ES/L3 and 99
for purified ES/L3. Using the ES/L3 purified antigen, it can be considered that the reactive sera, with compatible symptoms correspond to patients who are or were parasitized with Toxocara canis.