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Centro de Investigaciones Regionales Dr. Hideyo Noguchi, Universidad Autónoma de Yucatán

1999, Number 3

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Rev Biomed 1999; 10 (3)

Comparison of four antigens in immuno enzymatic assay (ELISA) for specific antibody determination in a serology of leptospirosis in dogs

Arias D, Arauz S, Stornelli A,Stanchi NO, Renner E, Martino PE, Gatti M

Language: Spanish
References: 21
Page: 167-171
PDF size: 26.55 Kb.


ABSTRACT

Introduction. Leptospirosis is a disease that frequently turns out sub diagnosed due to the lack an easily performed diagnostic test. Among the diagnostic techniques, the immuno enzymatic ones are among those of greater sensibility. The aim of this study is to develop the techniques of ELISA that allow for the diagnostic detection of Leptospirosis in a quick, simple and economic way.
Material and methods. Live cultivation of Leptospira interrogans sv icterohaemorrhagiae were used as antigen (Ag) with different treatment: total Ag, sonicated Ag, thermo resistant Ag (TR) and sonicated TR Ag. The test was carried out on 29 clinically suspicious canine sera of leptospirosis, diluted in PBS 1/8; using 3 negative and 3 positive controls. All sera were compared with Microscopic Agglutination Test (MAT) using as Ag the same strain used in ELISA.
Results. The results of this study show that out of 29 analyzed sera 10 (34%) were positive. All the positive control gave evident color development in the wells, with no change in the negatives ones. Correlation with the technique of MAT was +0.80 (sonicated TR) and +0.70 (TR) according to the antigen used.
Discussion. This quick technique (150 min) could be adapted to almost any laboratory and could be used as a diagnostic screening method of canine leptospirosis.


REFERENCES

  1. Baranton G. Control and prevention of Leptospirosis. Leptospirosis on the African Continent. Proceedings of the CEC/STD3 Research Meeting, Harare, Zimbabwe, The Netherlands: W. Terpstra; 1992. p. 17-20.

  2. Boerrigher G. Direct measurement of in vitro antibody production using and enzyme-linked immuno sorbent assay. J Immunol Methods 1983; 61:377-84.

  3. Tersptra W, Collares-Pereira M. ELISA for the diagnosis of Leptospirosis. Leptospirosis on the African Continent. Proceedings of the CEC/STD3 Research Meeting, Harare, Zimbabwe, The Netherlands: W.Terpstra; 1992. p. 17-20.

  4. Terpstra W, Ligthart GS, Schone GJ. Serodiagnosis of human Leptospirosis by enzyme-linked.immunosorbentassay (ELISA). Zbl Bakt I Abt Orig 1980; A247:400-5

  5. Banfi E, Cinco M, Delia S, Castagnari L, Vuello V, Mastorianni CM, et al. New trends in the rapid serodiagnosis of Leptospirosis. Zbl Bakt Mikrobiol Hyg I Abt Orig 1984; A257:503-7.

  6. Chapman AJ, Adler B, Faine S. Antigen recognized by the human immune response to infection with Leptospira interrogans serovar hardjo. J Med Microbiol 1988; 25:269-78.

  7. Chapman AJ, Adler B, Faine S. Antigens recognized by the human immune response to vaccinationwith a bivalent hardjo/pomona leptospiral vaccine. FEMS Microbiol Immunol 1990; 64:111-8.

  8. Adler B, Murphy AM, Lacardi SA, Faine S. Detection of specific anti-leptospiral immunoglobulin M and G in human serum by solid phase enzyme-linked immunosorbent assay. J Clin Microbiol 1980; 11:452-7.

  9. Faine S. Guidelines for the control of Leptospirosis. Geneva: WHO; 1982. p. 171. Publication nº 67.

  10. Kurstak E. Progress in enzyme immunoassays: production of reagents, control designs, and interpretation. Bull WHO 1985; 63:793-811.

  11. Ritaco V. Algunas sugerencias de interés práctico en ELISA. Uppsala: FAO/IAEA; 1987. p. 5.

  12. Kish L, Wiley J. EPI 6.0 Epidemiological Software. OMS. 1995.

  13. Zochowski WJ, Waitkins SA, Palmer MF. The use of ELISA in the diagnosis of human leptospirosis. Isr J Vet Med 1987; 43:330-9.

  14. Costa E. Estado imunólogico en Leptospirosis. Rev Inst A Lutz 1981; 41,2:93-100.

  15. Milner A. Enzyme linked immunosorbent assay for determining specific immunoglobulin M in infection caused by Leptospira interrogans sv hardjo. J Clin Microbiol 1985; 22:539-42.

  16. Watt G. The rapid diagnosis of Leptospirosis: a prospective comparison of the dog EIA, on the genus specific microscopic agglutination test at different stages of illness. J Infec Dis 1988; 157:840-2.

  17. Ellis W. Animal Leptospirosis. Constraints in diagnosis and research. Leptospirosis on the African Continent. Proceedings of the CEC/STD3 Research Meeting, Harare, Zimbabwe. The Netherlands: W. Terpstra; 1993. p. 17-20.

  18. Cousins D. Use of EIA in a serological survey of Leptospirosis in sheep. Aust Vet J 1986; 63:36-9.

  19. Gussenhoven G, Van Der Hoorn M, Goris M, Terpstra W, Hartskeerl R, Mol B, et al. LEPTO dipstick, a dipstick assay for detection of Leptospira-specific immunoglobulin M antibodies in human sera. J Clin Microbiol 1997; 35:92-7.

  20. Stanchi N, Pennimpede E, Gómez C. Serología de la Leptospirosis experimental en cobayos (Cavia porcellus). Fenómeno de zona. Acta Bioq Clin Latin 1991; 25:29-31.

  21. Takimoto T. Affinity constant of antileptospiral monoclonal antibody, numbers of antigenic determinants on leptospira, and their influence on the microscopic agglutination test and ELISA. Microbiol Immunobiol 1988; 32:775-84.




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