Evaluación de las pruebas DOT BLOT y aglutinación de látex para el diagnóstico de cisticercosis en Perú

Miranda-Ulloa, Eduardo; Sandoval-Ahumada, Roxana; Ayala, Eduardo; Vásquez-Ampuero, Juan Marco

doi:10.17843/rpmesp.2014.312.50

Cited by 3 publications

(5 citation statements)

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“…By that time, the adult worms are already installed in the bile conducts, generating processes of calcification, inflammation, and irreversible damage. In addition, neither of these two diagnostic methods reveals the parasite burden present in the infected individual (ruminants or humans), or if the infection is current or if it is an immunological trace with resolved infections, as in serology (Anuracpreeda et al 2016;Farrell et al 2015;Miranda-Ulloa et al 2014). This makes it necessary to develop sensitive and effective alternative techniques through which it may be possible to have early diagnosis of the infection (Valero et al 2009).…”

Section: Discussionmentioning

confidence: 99%

“…For Fasciola sp., no reports were found in the literature consulted on the development of rapid tests based on the latex agglutination reaction, although for other microorganisms (bacteria, protozoa) latex tests have proven highly useful with results [ 95% of specificity and sensitivity (Cuello-Pérez et al 2015;Darani et al 2010;Miranda-Ulloa et al 2014;Mohammadi et al 2013;Yu et al 2011).…”

Section: Discussionmentioning

confidence: 99%

“…antigens, through Dot Blot and latex agglutination tests. Another likely cause of crossed reaction may be due to the homology among the proteins of these parasites (Kindt et al 2007;Miranda-Ulloa et al 2014).…”

Section: Discussionmentioning

confidence: 99%

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Standardization of a latex agglutination test for coproantigen detection of Fasciola sp. in bovine cattle stool

et al. 2017

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Fasciolosis is a zoonotic parasitic disease, which affects humans and animals; diagnosed through noncommercial immunoassay tests that cannot be used on the field. Thereby, establishing the optimal conditions to develop a latex agglutination technique with IgG and IgM antibodies directed against excretion/secretion antigens of sp. is a priority. Latex particles were sensitized with IgG and IgM antibodies directed against excretion/secretion antigens of sp. The specificity of the antibodies was determined against antigens of different helminths and protozoa; the sensitivity and specificity of the test was evaluated against a previously standardized direct ELISA. The coupling rates of the IgG and IgM antibodies were 85.77 and 100%, respectively. The minimum detectable concentration of sp. excretion/secretion antigens, diluted in a phosphate-buffered saline, was 1.589 mg/mL(IgG) and 0.158 mg/mL(IgM) and for the antigens incorporated in the bovine cattle stool it was 3.178 mg/mL(IgG) and 1.589 mg/mL(IgM). The test showed crossed reaction against sp., and sp. antigens. Agreement of the IgG and IgM latex test against the ELISA test was of 78.78 and 96.96%, respectively; the specificity found was of 100% for both tests and sensitivity was 78.79% (IgG) and 96.97% (IgM). This work standardized the latex agglutination technique to detect sp. antigens in bovine cattle stool.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Standardization of a latex agglutination test for coproantigen detection of Fasciola sp. in bovine cattle stool

et al. 2017

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“…La técnica de Western Blot ha mejorado el diagnóstico de diversas enfermedades y tiene ventaja sobre otras técnicas serológicas existentes en el mercado que no son asequibles en zonas endémicas por el costo elevado, el requerimiento de un equipo adicional, y por el proceso y el tiempo de realización (22) .…”

Section: Introductionunclassified

“…No obstante, la prueba de ELISA se sigue empleando como técnica de tamizaje, pero requiere confirmación por Western Blot en caso el resultado sea reactivo; paso adicional que demanda mayor tiempo y costo (22) . El uso de Western Blot está sustentado por su eficiencia para el diagnóstico de la cisticercosis con una sensibilidad de 91%, determinada por Escalante, 98% por Tsang y 100% por Ayala (23) ; para hidatidosis entre 75 y 80% (24) y para fascioliasis humana 95% (25) ; todos con una especificidad del 100% (23)(24)(25) .…”

Section: Introductionunclassified

Rendimiento diagnóstico del Western Blot para detectar simultáneamente anticuerpos en pacientes con cisticercosis, hidatidosis y fascioliasis humana

Davelois¹,

Escalante²,

Jara

2016

Rev Peru Med Exp Salud Publica

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. To determine the diagnostic yield using western blotting to simultaneously detect antibodies in patients with human cysticercosis, hydatidosis, and human fascioliasis. Materials and methods . Cross-sectional study of diagnostic yield assessment. Excretory/secretory antigens were obtained from Taenia solium larvae, Echinococcus granulosus cysts, and the adult flukes of Fasciola hepática, which were then separated using the polyacrylamide gel electrophoresis technique, transferred, and attached to a nitrocellulose membrane to be probed with sera from the patient infected with the three parasites. The sensitivity of the technique was assessed using 300 individual serum samples, 60 pools of two parasites, and 20 pools of three parasites with 75 sera from patients with other parasites, 10 from patients with other diseases, and 15 from patients without parasites. Results . The technique revealed 13 glycoproteins (GP): GP 35, 31, 24, 23, 18, 17, 14, and 13 kDa for cysticercosis; GP 8, 16, and 21 kDa for hydatidosis; and GP 17 and 23 kDa for fascioliasis. The test detected the presence of antibodies with a sensitivity of 96% (95% confidence interval [CI] = 94.62-98.54%) in the detection of one or the thirteen bands, a specificity of 100% (95% CI = 99.50-100.00%); individually, there was a sensitivity for cysticercosis of 97% (95% CI = 93.16-100.00%), for hydatidosis of 94% (95% CI = 88.85-99.15%) and for fascioliasis of 96% (95% CI = 91.66-100.00%). Conclusions . Western blotting is effective in the simultaneous detection of antibodies in patients with human cysticercosis, hydatidosis, and fascioliasis, and it can be used as a diagnostic test to either rule out or confirm the presence of antibodies in endemic areas.

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Evaluación de electroinmunotransferencia utilizando antígeno mix nativo purificado de líquido de cisticerco de Taenia solium para el diagnóstico de cisticercosis humana

Ayala-Sulca

Miranda-Ulloa

2015

Rev Peru Med Exp Salud Publica

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Objetivos. 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 µg/mm. Por cromatografía de afinidad con lectina de lentejassefarosa 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.

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Evaluación de las pruebas DOT BLOT y aglutinación de látex para el diagnóstico de cisticercosis en Perú

Cited by 3 publications

References 10 publications

Standardization of a latex agglutination test for coproantigen detection of Fasciola sp. in bovine cattle stool

Standardization of a latex agglutination test for coproantigen detection of Fasciola sp. in bovine cattle stool

Rendimiento diagnóstico del Western Blot para detectar simultáneamente anticuerpos en pacientes con cisticercosis, hidatidosis y fascioliasis humana

Evaluación de electroinmunotransferencia utilizando antígeno mix nativo purificado de líquido de cisticerco de Taenia solium para el diagnóstico de cisticercosis humana

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