Resumo

FERRIOL, Xenia et al. Standardization and validation of immunoenzimatic assays for the quantitative determination of human IgG-class antibodies to leptospira canicola canicola, icterohaemorrhagiae copenhageni and pomona mozdok. Vaccimonitor [online]. 2001, vol.10, n.3, pp. 13-19. ISSN 1025-0298.

Indirect ELISAs for the quantitative determination of human IgG-class antibodies to leptospira were developed for the evaluation of the immune response elicited by vax-SPIRAL. Whole cells of Leptospira interrogans, serovars canicola canicola, icterohaemorrhagiae copenhageni and pomona mozdok were used as capture antigens. The formaldehydeinactivated cells were dried at 33 °C between 16-20 hours on ELISA plates. The units of the standard serum were assigned according the inverted microagglutination tittles, and were 31, 12 and 58 U/mL respectively. An anti-human IgG class / HRPO conjugate was employed for the detection of antibodies to leptospira, then the chromogenic substrate (H2O2-orthophenilendiamine) is hydrolyzed by the enzyme of the conjugate, and the intensity of the color is proportional to the antibody concentration in the sample. The intra and interassay precision studies achieved variation coefficients below 10%. The deviations of the recovery and linearity tests were below 10% also. The control serum was located according the samples. The sensitivity was 100% for all serovars, and the specificity was 93,33% for the serovars canicola canicola and icterohaemorrhagiae copenhageni, and 100 % for pomona mozdok. The detection limits were 0.478, 0,127 and 0,632 U/mL respectively.

Palavras-chave : Leptospira interrogans; validation; ELISA; vax-SPIRAL.

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