Early Diagnostic Test for hydatid cyst in high mountain alpacas
DOI:
https://doi.org/10.54943/rcsxxi.v4i2.566Keywords:
antigen, polyclonal antibodies, characterization, hydatid disease, protoscólexAbstract
The purpose of the research work was to develop an early diagnostic test for hydatid cyst in alpacas using the Enzyme Linked Immunosorbent Assay (ELISA) technique. Vertical electrophoresis methods, serological gel immunodiffusion methods and the ELISA technique were used as a diagnostic test. For this process, 8 alpaca hydatid cyst samples were accessed, then the viability of the cysts was determined by observing the protoscolysis and scabs of Echinococcus granulosus under a microscope. Then we continued with the characterization of antigenic proteins, recognizing the Ag5 antigen (subunits 67.72 and 55.44 KDa) and the AgB antigen (subunits 24.57, 16.79 and 12.10 KDa). To determine polyclonal anti-hydatid cyst antibodies, alpaca antigens were inoculated in 2 rabbits and verification was carried out by observing precipitation bands in the antigen-antibody reaction at a dilution of 1/16 and concentration of 1.26ug/µL. by the immunodiffusion method. In the end, the ELISA technique was used to determine antibodies against the hydatid cyst, obtaining a low sensitivity of 13.5% and a specificity of 57.1%, due to cross-reaction with other parasites due to working with total antigens. Concluding, that the Ag5 and AgB of the fluid of hydatid cysts of alpacas were characterized, polyclonal anti-hydatid cyst antibodies were obtained in rabbits and a low sensitivity and specificity was achieved in the diagnostic test.
Downloads
References
Almeida F., Rodrigues-Silva R., Neves R., Romaní E. and Machado-Silva J. 2007. Intraespecific variation of Echinococcus granulosus in livestock from Peru. Veterinary Parasitology. 147: 50 – 58.
Alho, A. M., Dias, M. C., Cardo, M., Aguiar, P., & de Carvalho, L. M. (2023). The Evolution of Cystic Echinococcosis in Humans and Ruminants in Portugal-A One Health Approach. Veterinary sciences, 10(9), 584
Craig, P. S., Gasser, R. B., Parada, L., Cabrera, P., Parietti, S., Borgues, C., & Paolillo, E. (1995). Diagnosis of canine echinococcosis: comparison of coproantigen and serum antibody tests with arecoline purgation in Uruguay. Veterinary parasitology, 56(4), 293-301.
Daniel, WW. (2004). Bioestadística base para el análisis de las ciencias de la salud. Editorial Limusa Wiley WAYNE W. DANIEL.
Daeki, AO. (2000). IgG-subclass antibody responses and the natural history of hepatic cystic echinococcosis in asymptomatic patients. Ann Trop Med Parasitol; 94: 319-328.
Di Felice, G., Pini, C., Afferni, C., Vicari, G. (1986). Purification and partial characterization of the major antigen of Echinococcus granulosus (antigen 5) with monoclonal antibodies. Parasitologia Molecular y Bioquimica. 20 (2). 133-142.
Dunne, DW.; Grabowska, AM.; Fulford, AJ. et al. (1988). Human antibody responses to S. mansoni: the influence of epitopes shared between different lifecycle stages on the response to the schistosomulum. Europ. J. Immun., 18: 123-131.
Guisantes, J. A., & Yarzábal, L. A. (1975). Diagnóstico de la hidatidósis mediante la doble difusión en gel. Boletín de la Oficina Sanitaria Panamericana (OSP); 78 (3), mar. 1975.
Grimm, F. (1998). Analysis of specific immunoglobulin G subclass antibodies for serological diagnosis of echinococcosis by standard enzyme-linked immunosorbent assay. Clin Diag Labor Immunol; 5: 613-616.
Hajjafari, A., Sadr, S., Santucciu, C., Masala, G., Bayat, M., Lotfalizadeh, N., Borji, H., Partovi Moghaddam, S., y Hajjafari, K. (2024). Avances en la detección de la equinococosis quística en huéspedes intermediarios y nuevas herramientas de diagnóstico: una revisión de la literatura. Ciencias Veterinarias , 11 (6), 227.
Irabuena, O., Nieto, A., Ferreira, A. M., Battistoni, J., & Ferragut, G. (2000). Characterization and optimization of bovine Echinococcus granulosus cyst fluid to be used in immunodiagnosis of hydatid disease by ELISA. Revista do Instituto de Medicina Tropical de São Paulo, 42, 255-262.
Li, J., Zhang, WB, Wilson, M., Ito, A., y McManus, DP (2003). Un nuevo antígeno recombinante para el inmunodiagnóstico de la equinococosis quística humana. The Journal of infectious diseases , 188 (12), 1951-1960.
Lightowlers, MW., Liu, DY., Haralambous, A., Rickard, MD. (1989). Subunit composition and specificity of the major cyst fluid antígena of Echinococcus granulosus. Molecular and Biochemical Parasitology. 37(2): 171-182.
Lomonte, B. (2007). Manual de métodos inmunológicos. Consultado el 15 de octubre del 2014, de http://repositorio.ucr.ac.cr/handle/10669/9244.
Miranda, E., Velarde, F., Somocurcio, J., Ayala, E. (2010). Evaluación de dos pruebas de Inmunoblot con antígeno hidatídico de caprino y ovino para el diagnóstico de equinococosis humana. Rev Peru Med Exp Salud Publica. 2010; 27(2): 209-14.
Moro, P., Schantz, P. (2009). Echinococcosis: a review. Int J Infect Dis; 13(2): 125-133.
Mamuti, W., Yamasaki, H., Sako, Y., Nakaya, K., Nakao, M., Lightower’s, M.W, Ito A. (2002). Usefulness of Hydatid Cyst Fluid of Echinococcus granulosus Developed in Mice with Secondary Infection for Serodiagnosis of Cystic Echinococcosis in Humans. Clin Vaccine Immunol. 9(3): 573-576
Naquira, C. (1994). Hydatidosis situation in Perú. In Proc.Scientific Working group in the advances in the prevention, control and treatment of hydatidosis.
Oriol, R., Williams, JF., Perez-Esandi, MV., Oriol, C. (1971). Purification of lipoprotein antigens of Echinococcus granulosus from sheep hydatid fluid. The American Society of Tropical Medicine and Hygiene. 20(4): 569-574.
Pérez Esandi, M. V. (1970). Aislamiento y caracterización de anticuerpos en sueros de humanos infectados con Echinococcus granulosus. Boletín de la Oficina Sanitaria Panamericana (OSP); 69 (1), jul. 1970.
Peruzzu, A., Mastrandrea, S., Fancellu, A., Bonelli, P., Muehlethaler, K., Masala, G., & Santucciu, C. (2022). Comparison and evaluation of analytic and diagnostic performances of four commercial kits for the detection of antibodies against Echinococcus granulosus and multilocularis in human sera. Comparative immunology, microbiology and infectious diseases, 86, 101816,
Rigano, R., Buttari, B., Profumo, E., (2007). Echinococcus granulosus antigen B impairs human dendritic cell differentiation and polarizes immature dendritic cell maturation towards a Th2 cell response. Infect. Immun. 75, 1667–78.
Rojas, M. (1990). Parasitismo de los Rumiantes Domésticos. Terapia, Prevención y Modelos para su aprendizaje. Editorial Mijosa. Primera edición. Lima - Perú. 335 - 343.
Sánchez, EL., Naquira, CG., Vega, ES., Miranda, EF., Quispe, WM., Ayala, ER. (2010). Manual de Procedimiento para el Diagnostico Serológico de las Zoonosis Parasitarias. Perú. Segunda edición. Catalogación hecha por el Centro de Documentación e Información del INS.
Sánchez, F. (1992). Caracterización, Purificación y localización Inmunohistoquímica de los antígenos mayoritarios de Echinococcus granulosus Antígeno 5 y Antígeno B. (Tesis de pos grado). Universidad Autónoma de Barcelona.
Schantz, P.M., Chai, J., Craig, P.S., Eckert, J., Jenkins, D.J., Macpherson, C.N.L., Thakur, A., (1995). Epidemiology and control of hydatid disease. In: Thompson, R.C.A., Lymbery, A.J. (Eds.), Echinococ- cus and Hydatid Disease. CAB International, Wallingford, UK.
Serra, E., Masu, G., Chisu, V., Cappai, S., Masala, G., Loi, F., & Piseddu, T. (2022). Contaminación ambiental por huevos de Echinococcus spp. como riesgo para la salud humana en granjas educativas de Cerdeña, Italia. Ciencias veterinarias , 9 (3), 143.
Siracusano, A. (1991). Detection of antibodies against Echinococcus granulosus major antigens and their subunits by immunoblotting. Transmit Res Soc Trop Med Hyg; 85 (2): 239-243.
Vargas, D., Bonet, R., Jofré, C., Campano, S. (2005). Evaluación del líquido hidatídico como sustrato de sensibilización en la obtención de anticuerpos policlonales de uso en ELISA coproantigénica. Parasitol Latinoam: 60 (1-2). 93-96.
Vildózola, H., Espinoza, I., Roldan, WH. (2012). Estandarización de una prueba de ELISA para detectar anticuerpos IgE en pacientes con equinococosis quística y su utilidad en el diagnóstico y seguimiento de pacientes tratados con albendazol. An. Fac. med.73 (1).
Zhang W, Li J, McManus DP.( 2003) Concepts in immunology and diagnosis of hydatid disease. Clin Microbiol Rev; 16:18–36.
Widdicombe J, Basáñez MG, Entezami M, Jackson D, Larrieu E, Prada JM (2022) La evaluación económica de las estrategias de control de la equinococosis quística centradas en los huéspedes zoonóticos: una revisión de alcance. PLoS Negl Trop Dis 16(7): e0010568
Published
How to Cite
-
Abstract385
-
PDF (Español (España))123
-
HTML (Español (España))15
Issue
Section
License
Copyright (c) 2025 Nicasio Valencia Mamani, Luz Marina Vilcapaza Quispe, Blas Reymundo Condor, Wiliam Quispe Paredes, Epifanio Lizana -Hilario, Catherine Charlotte Carrasco Canepa, Victor Carhuapoma-Delacruz
This work is licensed under a Creative Commons Attribution 4.0 International License.
