Standardization of the dot enzyme-lynked immunosorbent assay (dot-ELISA) for experimental plague

Memórias do Instituto Oswaldo Cruz

Silvia M. L. Montenegro²  Alzira M. P. De Almeida²  Luiz B. Carvalho Júnior¹ 

[1] ,FIOCRUZ Centro de Pesquisas Aggeu Magalhães Recife,Brasil

关键词: dot-ELISA;    dacron;    Yersinia pestis;    solid-phase;   

DOI  :  10.1590/S0074-02761993000100018

来源: SciELO

【 摘 要 】

A dot enzyme linked immunosorbent assay (dot-ELISA) was previously developed to detect specific antibodies in rabbits sera immunized against FIA protein obtained from Yersina pestis. This antigen was covalently linked onto the surface of dacron (polyethyleneterephthalate). Here, standard conditions are described for the optimization of this procedure: an amount of 20 ng of FIA protein was fixed onto dacron; anti-rabbit IgG peroxidase conjugate diluted 1:8,000 and 30% non-fat instant milk as blocking substance were used throughout the method. This procedure was compared with that employing nitrocellulose as solid-phase which showed to be more sensitive. However, the method based on dacron did not show false positive reactions against non-immunized rabbits sera at low antigen amount and diluted anti-IgG peroxidase conjugate.

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