medigraphic.com
Revista Latinoamericana de Microbiología

2000, Number 2

<< Back Next >>

Microbiología 2000; 42 (2)

Experimental swine Trichinellosis: Use of Dot-ELISA and western blot with excretion/secretion antigens (ES) from infective larvae to detect anti-Trichinella spiralis antibodies.

Aguilar FBR, Bautista-Garfias CR, Rojas J, de Nova ME, Rodríguez OI, Martinez-Gómez F

Language: English
References: 23
Page: 57-62
PDF size: 152.82 Kb.


ABSTRACT

Excretion/secretion antigens from Trichinella spiralis infective larvae were evaluated in Dot-ELISA and Western blot techniques for the serologic diagnosis of trichinellosis in experimentally infected swine. Three York female pigs were infected with 10,000 T. spiralis infective larvae/animal. After infection, weekly blood samples, during 10 weeks, were taken from them to obtain sera. At this time muscle tissue samples were collected from tongue and diaphragm and subjected to artificial digestion to determine the number of larvae per gram of muscle tissue (LPGMT). Animals showed a mean of 10 LPGMT. Positive anti-T. spiralis antibody titers (threshold of positivity 1:160) were determined in sera five weeks after infection by Dot-ELISA. When the Western blot method was used a pattern of seven antigenic components of 229, 108, 64, 45, 41, 40 and 33-38.5 kDa were recognized by swine sera at week five after infection. Sera from Ascaris suum infected-swine were negative to anti-Trichinella antibodies when analyzed by Dot-ELISA and Western blot. Both analysis were similar in discriminating positive from non-positive sera to anti-T. spiralis antibodies and showed a sensitivity and specificity of 100%. For this reasons and for its easily to carry out, it is suggested to use Dot-ELISA for the serologic diagnosis of swine trichinellosis.


REFERENCES

  1. Arriaga, C., L Yepez-Mulia, A. Morilla. and M. G. Ortega-Pierres. 1995. Detection of circulating Trichinella spiralis muscle larva antigens in serum samples of experimentally and naturally infected swine. Vet. Parasitol 58:319-326.

  2. Correa, D., L. De La Rosa, M. G. Ortega-Pierres. 1997. Trichinellosis: Human epidemiological data in Mexico from 1939 to 1995. In: G. Ortega-Pierres, H.R. Ga mble, F. van Knapen, D. Wakelin (eds.) Trichinellosis ICT9. Centro de Investigación y Estudios Avanzados del Instituto Politécnico Nacional, México, D. F. México. pp. 519-523.

  3. Despommier, D.D., A. M. Gold, and W. S. Buck. 1990. Partial characterization of two antigens secreted by L1 larva of Trichinella spiralis. Mol. Biochem. Parasitol. 41:187-196.

  4. Gamble , H. R., W. R. Anderson, C. E. Graham and K. D. Murrell. 1983. Diagnosis of swine trichinosis by enzyme-linked immunosorbent assay (ELISA) using an excretory-secretory antigen. Vet. Parasitol. 13:349-361.

  5. Gamble , H. R., D. Rapic, A. Marinculic, and K. D. Murrell. 1988. Evaluation of excretory-secretory antigens for the serodiagnosis of swine trichinellosis. Vet. Parasitol. 30:131-137.

  6. Gamble , H. R., N. Winsnewski, D. L. and D. L. Wason. 1997. Diagnosis of trichinellosis in swine by enzyme immunoassay, using a synthetic glycan antigen. Am. J. Vet. Res. 58:1417-1421.

  7. Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-684.

  8. Lowry, O. H., N. J. Rosebrough, A. L. Farr, and R. J. Randall. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265-275.

  9. Martínez-Marañón, R. 1985. ¿Está aumentando la triquinosis en México? ¿Podría ser una consecuencia inesperada de nuestro "desarrollo"? Sal. Publ. Mex. 27:40-51.

  10. NCCLS 1993. Procedures for the recovery and identification of parasites from the intestinal tract, proposed guideline. NCCLS Document M28-P Vol. 13 No. 20.

  11. Nockler, K., W. P. Voigt, D. Protz, and K. Ziedler. 1995. Diagnosis of trichinellosis in living pigs using indirect ELISA. Berl. Munch. Tierarztl. Wochenschr. 108:167-174.

  12. Pappas, G. M., R. Hajkowski, L. T. Cannon, and W. T. Hockmeyer. 1984. Dot enzyme-linked immunosorbent assay (Dot-ELISA): Comparis on with standard ELISA and complement fixation assays for the diagnosis of human visceral leishmaniasis. Vet. Parasitol. 14:239-249.

  13. Peña-Piña, A., C. R. Bautista Garfias, M. CastillaGarcia. 1988. Comparación de la prueba de inmunoensayo en capa delgada (ICD) con la técnica de hemoaglutinación pasiva (HP) para la detección de anticuerpos anti-Trichinella spiralis en roedores. Vet. Méx. 19:321-327.

  14. Ramírez-Valenzuela, M. 1985. La triquinelosis en México. Un estudio epidemiológico retrospectivo. En: A. Morilla, P. Correa, A. Stephano (eds.) Avances en Enfermedades del Cerdo. Ediciones de la Asociación Mexicana de Veterinarios Especialistas en Cerdos, A. C. México, D. F. México. pp. 557-573.

  15. Rohrbacher, G. H. 1957. Observations on the survival in vitro of bacteria-free adult common liver flukes, Fasciola hepatica. J. Parasitol. 43:9-18.

  16. Ruitenberg, E. J., P. A. Steeerenberg, B. J. M. Brossi, and J. Buys. 1974. Serodiagnosis of Trichinella spiralis infections in pigs by enzyme-linked immunosorbent assays. Bull. W.H.O. 51:108-109.

  17. Ruitenberg, E. J., F. Van Knapen, and A. Elgersma. 1983. Surveillance in swine by immunodiagnostic methods. In. W. C. Campbell (ed.) Trichinella and Trichinosis. Plenum Press. New York. pp. 529-550.

  18. Seawright, G. L., D. D. Despommier, W. Zimmerman, and R. S. Isenstein. 1983. Enzyme immunoassay for swine trichinellosis using antigens purified by immunoaffinity chromatography. Am. J. Trop. Med. Hyg., 32:1275-1284.

  19. Shin, S. S., F. Elvinger, A. K. Prestwood and J. R. Cole . 1997. Exposure of swine to Trichinella spiralis antigen as determined by consecutive ELISAs and western blot. J. Parasitol. 83:430-433.

  20. Su, X. and A. K. Prestwood. 1991. A dot-ELISA mimicry Western blot test for the detection of swine trichinellosis. J. Parasitol. 77:76-82.

  21. Van Knapen, F., J. H. Franchimont, E. J. Ruitenberg, B. Baldelli, J. Bradley, T. E. Gibson, C. Gottal, S. A. Henriksen, S. A., G. Kohler, N. Skovgaard, C. Soule, and S. M. Taylor. 1980. Comparison of the enzymelinked immunosorbent assay (ELISA) with three other methods for the detection of Trichinella spiralis infections in pigs. Vet. Parasitol. 7:109-121.

  22. Venturiello, S. M., G. J. BEN, S. N. Constantino, S. L. Malmassari, G. G. Nunez, R. L. Veneroni, and M. J. Traversa. 1998. Diagnosis of porcine trichinellosis: parasitological and immunoserological tests in pigs from endemic areas of Argentina. Vet. Parasitol. 74: 215-228.

  23. 23.Weiger, L. H., C. G. Denksen and F. Van Knapen. 1992. Identification of diagnostic antigens from Trichinella spiralis. Parasitol. Res. 78:112-119.




2020     |     www.medigraphic.com