Evaluation of an in-house enzyme linked immuonosorbent assay (ELISA) for detection of anti – Fasciola gigantica antibodies in sheep

Document Type : Research Articles

Authors

• Mohammad Hossein Razi Jalali ¹
• Hossein Hamidinejat ²
• Massoud Ghorbanpoor ¹
• Saleh Esmaeelzadeh ¹
• Abbas Sohbati ³

¹ Departments of Pathobiology, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz

² Department of Clinical Sciences, Faculty of Veterinary Medicine, Shahid Chamran University of Ahvaz, Ahvaz

³ Graduated of parasitology, Faculty of veterinary medicine , Shahid Chamran u nivercity , Ahvaz

10.22067/veterinary.v4i2.16169

Abstract

Tropical fasciolosis caused by infection with Fasciola gigantica is regarded as one of the most important single helminthes infections of ruminants in Asia and Africa. Fasciolosis diagnosis, due to low sensitivity of coprological diagnostic method has been challenging for a long period. To evaluate the usefulness of ELISA for diagnosis of anti – Fasciola gigantica antibodies in parasitized sheep, crude, as well as excretory-secretory (E/S) antigens were examined with 50 positive and 45 negative serum samples. Results were compared with liver inspection in slaughterhouse. The cut off values were determined in 0.469 and 0.516 for crude and E/S antigen respectively. We found 82% of sensitivity, 95.6 of specificity, 95.3% positive predictive value and 83 % negative predictive value using crude antigen. We have also found 92% of sensitivity, 93% specificity, 93.8% positive predictive value and 91.4 % negative predictive value using E/S antigen. We concluded that ELISA, specially using E/S antigen, is a good tool for serodiagnosis of Fasciola gigantica infection in sheep herds.

Keywords

• Fasciola gigantic
• ELISA
• Sheep

20.1001.1.2008465.2012.4.2.3.5

Main Subjects

• Enzymology
• Immunology

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References

Anderson, N., Luong, T.T. Vo, N.G. Bui, K.L. Smooker, P.M, Spithill, T.W. (1999) The sensitivity and specificity of two methods for detecting Fasciola infections in cattle. Veterinary Parasitology, 83, 15–24.

 

Boray, J.C. (1997) Immunology, Pathobiology and Control of Fasciolosis. MSD AGVET, Rahway, New Jersey, USA.

 

Castro, E., Freyre A., Hernandez, Z. (2000) Serological responses of cattle after treatment and during natural re-infection with Fasciola hepatica measured using a dot-ELISA system, Veterinary Parasitology, 90, 201–208.

 

Charlier, J., De Meulemeester, L. Claerebout Williams, E.D., Vercruysse, J. ( 2008) Qualitative and quantitative evaluation of coprological and serological techniques for the diagnosis of fasciolosis in cattle. Veterinary Parasitology, 153, 44–51.

 

Fagbemi, B.O., Obarisiagbon, I.O. ( 1990) Comparative evaluation of the enzyme linked immunosorbent assay in the diagnosis of natural Fasciola gigantica infection in cattle. Veterinary Quarterly, 12, 35-8.

 

Hillyer, G.V., Sanchez, Z., De Leon, D.(1985) Immunodiagnosis of bovine fascioliasis by enzyme-linkedimmunosorbent assay and immunoprecipitation methods. Journal of Parasitology, 71, 449-54.

 

Hillyer, G.V. (1986) Fasciolosis and fasciolopsiasis. In: Balows, A., Hausler, W.J., Ohashi, M. and Turano, A. (Eds): Laboratory Diagnosis of Infectious Disease. Springer-Verlag, Berlin, 90, 856-862.

 

Hillyer, G.V., Soler de Galanaes, M. , Buchon, P., Bjorland, J. (1996) Herd evaluation by enzyme-linked immunosorbent assay for the determination of Fasciola hepatica infection in sheep and cattle from the Altiplano of Bolivia. Veterinary Parasitology, 61, 211-220.

 

Kralova-Hramadova, I., Spakulova, M. Horackova, E. Turcekova, L. Novobilsky, A. Beck, R. Koudela, B. Marinculic, A. Rajsky, D., Pybus, M.(2008) Sequences analysis of ribosomal and mitochondrial genes of the giant liver fluke Fascioloides magna (Trematoda: Fasciolidae): intraspecific variation and differentiation from Fasciola hepatica. Journal of Parasitology, 98, 58-67.

 

Leeflang, M.G., Bossuyt, P.M.M. (2005) Test accuracy is likely to vary depending on the population it is used in. Veterinary Parasitology 134, 189.

 

Leevieux, D., Levieux, A. Mage, C., Venien, A.(1992) Early immunodiagnosis of bovine fascioliasis using the specific antigen f2 in a passive hemagglutination test. Veterinary Parasitology, 44, 77- 86.

 

Lowry, O.H., Rosenbrough, N.J. . Fair, A.L., Randall, R. (1951) Protein measurement with The Folin phenol reagent. Journal of Biological Chemistry., 193, 265–275.

 

Makay, O., Gurcu, B. Caliskan, C. Nart, D. Tuncyurek, M., Korkut, M.(2007) Ectopic fascioliasis mimicking a colon tumor. World Journal of Gastroenterology, 13, 2633-2635.

 

Oldham, G., Williams, L. (1985) Cell mediated immunity to liver fluke antigens during Experimental Fasciola hepatica infection of cattle. Parasite Immunology., 7, 503-516.

 

Rivera-Morero, C.A., Santiago, N., Hillyer, G.V. (1988) Evaluation of immunodiagnostic antigens in the excretory-secretory product of Fasciola hepatica. Journal of Parasitology, 74, 646-652.

 

Rokni, M.B., Samani, A., Massoud J., Babaei Nasr, M. (2006) Evaluation of Dot-ELISA Method Using Excretory-Secretary Antigens of Fasciola hepatica in Laboratory Diagnosis of Human Fasciolosis. Iran. Journal of Parasitology, 1, 26-30.

Sanchez Andrade, R., Paz Silva, A., Suarez, J., Pandero, R., DiezBanoy, P., Morrondo, P.(2000) Use of a sandwich- enzyme- linked immunosorbent assay (SEA) for the diagnosis of natural Fasciola hepatica infection in cattle from Galicia (NW Spain). Veterinary Parasitology, 93, 39-46.

 

Santiago, N., Hillyer, G.V. (1988) Antibody profiles by EITB and ELISA of cattle and sheep infected with Fasciola hepatica. Journal of Parasitology, 74, 810-8.

 

Simsek, S., Koroglu, E., Utuk, A.E., Altay, K. (2006) Use of indirect excretory/secretory enzyme-linked immunosorbent assay (ES-ELISA) for the diagnosis of natural Fasciola hepatica infection in eosinophilic and noneosinophilic cattle from Eastern Turkey. Turkish Journal of Veterinary And Animal Sciences, 30, 411–415.

 

Sukapesna, V., Tuntasuvan,D., Sarataphan, N., Imsub K. (1994) Economic impact of fascioliasis in buffalo production. The Thai Journal of Veterinary Medicine, 45, 45-52.

 

Viyanant, V., Krailas, D., Sobhon, P., Upatham, E.S., Kusamran, T., Chompoochan, Thammasart, S., Prasittirat, P. (1997). Diagnosis of cattle fasciolosis by the detection of a circulating antigen using a monoclonal antibody. Asian Pacific Journal of Allergy & Immunology, 15, 153-159.

 

Whiting, P., Rutjes, A.W., Reitsma, J.B., Glas, A.S.,Bossuyt, P.M., Kleijnen, J. (2004)

Sources of variation and bias in studies of diagnostic accuracy: a systematic review. Annals of Internal Medicine, 140, 189–202.

 

Zimmerman, G.L., Kerkvliet, N.Y., Brauner J.A., Cerro, J.E. (1983) Modulation of the host immune response by Fasciola hepatica: response of peripherallymphocyte to mitogens during liver fluke infection of sheep. Journal of Parasitology, 69, 473–477.