ABSTRACT
OBJECTIVES: Evaluate the efficiency of the enzyme-linked immunoelectrotransfer blot test (EITB) for the detection of human cysticercosis using purified native antigen mix from cysticercus fluid of Taenia solium. MATERIAL AND METHODS: Observational study of the evaluation of the diagnostic test. The purified native antigen mix was extracted from the fluid of cysticercus from naturally parasitized pigs from areas considered endemic. Four purification methods were evaluated and one was selected: ammonium sulfate, lentil lectin-Sepharose, Sephadex G-75 and Electro-elution. The sensitivity of the purified antigen was determined with EITB and was evaluated with 50 sera positive for cysticercosis and the specificity with 50 sera negative for cysticercosis (20 free of parasitic infection and 30 positive for different parasites). RESULTS: The optimal concentration of the purified native antigen mix to prepare the EITB strips was 0.82 µg/mm. By affinity chromatography with Lentil Lectin- Sepharose we visualized and purified 8 specific antigenic glycoproteins, their relative masses being: 13, 14, 17, 18, 23, 24, 31 and 35 kDa. The purified antigens obtained by the other methods were not evaluated by EITB because they did not correspond to the specific antigenic proteins considered between 13 and 35 kDa. The EITB test using purified native antigen mix showed 100% sensitivity and 100% specificity. CONCLUSIONS: The purified antigen native mix improved the diagnostic efficiency of the EITB test. We recommend the preparation of an in-house kit and field validation so that it can be transferred and implemented in laboratories in endemic areas of Peru.
Subject(s)
Antigens, Helminth , Cysticercosis/diagnosis , Enzyme-Linked Immunosorbent Assay , Taenia solium , Animals , Humans , Peru , SwineABSTRACT
Evaluar la eficacia de la prueba electroinmunotransferencia (EITB) para la detección de cisticercosis humana utilizando antígeno mix nativo purificado de líquido de cisticerco de Taenia solium. Material y métodos. Estudio observacional de evaluación de prueba diagnóstica. El antígeno mix nativo purificado se extrajo del líquido de cisticerco de cerdos naturalmente parasitados de áreas consideradas endémicas, se evaluó y seleccionó uno de los cuatro métodos de purificación: sulfato de amonio, lecitina de lentejas-sefarosa, Sephadex G-75 y electro-elución. La sensibilidad del antígeno purificado se determinó con EITB y fue evaluada con 50 sueros positivos a cisticercosis y la especificidad con 50 sueros negativos a cisticercosis (20 libres de infección parasitaria y 30 positivos a diferentes parasitosis). Resultados. La concentración óptima del antígeno mix nativo purificado para preparar las tiras EITB fue de 0,82 ug/mm. Por cromatografía de afinidad con lectina de lentejas-sefarosa se visualizaron y purificaron ocho glicoproteínas antigénicas específicas, siendo sus masas relativas de: 13, 14, 17, 18, 23, 24, 31 y 35 kDa; los antígenos purificados obtenidos por los otros métodos no fueron evaluados por EITB, porque no correspondieron a las proteínas antigénicas específicas consideradas entre 13 y 35 KDa. La prueba de EITB utilizando antígeno mix nativo purificado presentó 100% de sensibilidad y 100% de especificidad. Conclusiones. El antígeno mix nativo purificado mejora la eficiencia diagnóstica de la prueba de EITB. Recomendamos la preparación de un kit in house y su validación en campo para que pueda transferirse e implementarse en laboratorios de zonas endémicas del Perú...
Evaluate the efficiency of the enzyme-linked immunoelectrotransfer blot test (EITB) for the detection of human cysticercosis using purified native antigen mix from cysticercus fluid of Taenia solium. Material and methods. Observational study of the evaluation of the diagnostic test. The purified native antigen mix was extracted from the fluid of cysticercus from naturally parasitized pigs from areas considered endemic. Four purification methods were evaluated and one was selected: ammonium sulfate, lentil lectin-Sepharose, Sephadex G-75 and Electro-elution. The sensitivity of the purified antigen was determined with EITB and was evaluated with 50 sera positive for cysticercosis and the specificity with 50 sera negative for cysticercosis (20 free of parasitic infection and 30 positive for different parasites). Results. The optimal concentration of the purified native antigen mix to prepare the EITB strips was 0.82 ug/mm. By affinity chromatography with Lentil Lectin-Sepharose we visualized and purified 8 specific antigenic glycoproteins, their relative masses being: 13, 14, 17, 18, 23, 24, 31 and 35 kDa. The purified antigens obtained by the other methods were not evaluated by EITB because they did not correspond to the specific antigenic proteins considered between 13 and 35 kDa. The EITB test using purified native antigen mix showed 100% sensitivity and 100% specificity. Conclusions. The purified antigen native mix improved the diagnostic efficiency of the EITB test. We recommend the preparation of an in-house kit and field validation so that it can be transferred and implemented in laboratories in endemic areas of Peru...
Subject(s)
Humans , Antigens , Cysticercosis/diagnosis , Taenia solium , Observational Studies as TopicABSTRACT
El presente manual uniformiza los procedimientos de diagnóstico serológico de las principales zoonosis parasitarias estandarizados en el Laboratorio de Zoonosis Parasitarias del Centro Nacional de Salud Pública del Instituto Nacional de Salud. En esta segunda edición se ha considerado, además de las técnicas ya existentes, la prueba de ELISA y la técnica de aglutinación de látex para el diagnóstico de la hidatidosis humana
Subject(s)
Serologic Tests , Zoonoses , Zoonoses/parasitology , PeruABSTRACT
El presente manual uniformiza los procedimientos de diagnóstico serológico de las principales zoonosis parasitarias estandarizados en el Laboratorio de Zoonosis Parasitarias del Centro Nacional de Salud Pública del Instituto Nacional de Salud. En esta segunda edición se ha considerado, además de las técnicas ya existentes, la prueba de ELISA y la técnica de aglutinación de látex para el diagnóstico de la hidatidosis humana(AU)
