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

Autor: Silvia M. L. Montenegro, Alzira M. P. de Almeida, Luiz B. Carvalho Júnior
Jazyk: angličtina
Rok vydání: 1993
Předmět:
Zdroj: Memorias do Instituto Oswaldo Cruz, Vol 88, Iss 1, Pp 119-123 (1993)
Druh dokumentu: article
ISSN: 0074-0276
1678-8060
DOI: 10.1590/S0074-02761993000100018
Popis: 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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