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

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

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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Main Authors: Montenegro,Silvia M. L., Almeida,Alzira M. P. de, Carvalho Júnior,Luiz B.
Format: Digital revista
Language:English
Published: Instituto Oswaldo Cruz, Ministério da Saúde 1993
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02761993000100018
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spelling oai:scielo:S0074-027619930001000182009-06-01Standardization of the dot enzyme-lynked immunosorbent assay (dot-ELISA) for experimental plagueMontenegro,Silvia M. L.Almeida,Alzira M. P. deCarvalho Júnior,Luiz B. dot-ELISA dacron Yersinia pestis solid-phase 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.info:eu-repo/semantics/openAccessInstituto Oswaldo Cruz, Ministério da SaúdeMemórias do Instituto Oswaldo Cruz v.88 n.1 19931993-03-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02761993000100018en10.1590/S0074-02761993000100018
institution SCIELO
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country Brasil
countrycode BR
component Revista
access En linea
databasecode rev-scielo-br
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region America del Sur
libraryname SciELO
language English
format Digital
author Montenegro,Silvia M. L.
Almeida,Alzira M. P. de
Carvalho Júnior,Luiz B.
spellingShingle Montenegro,Silvia M. L.
Almeida,Alzira M. P. de
Carvalho Júnior,Luiz B.
Standardization of the dot enzyme-lynked immunosorbent assay (dot-ELISA) for experimental plague
author_facet Montenegro,Silvia M. L.
Almeida,Alzira M. P. de
Carvalho Júnior,Luiz B.
author_sort Montenegro,Silvia M. L.
title Standardization of the dot enzyme-lynked immunosorbent assay (dot-ELISA) for experimental plague
title_short Standardization of the dot enzyme-lynked immunosorbent assay (dot-ELISA) for experimental plague
title_full Standardization of the dot enzyme-lynked immunosorbent assay (dot-ELISA) for experimental plague
title_fullStr Standardization of the dot enzyme-lynked immunosorbent assay (dot-ELISA) for experimental plague
title_full_unstemmed Standardization of the dot enzyme-lynked immunosorbent assay (dot-ELISA) for experimental plague
title_sort standardization of the dot enzyme-lynked immunosorbent assay (dot-elisa) for experimental plague
description 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.
publisher Instituto Oswaldo Cruz, Ministério da Saúde
publishDate 1993
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02761993000100018
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AT carvalhojuniorluizb standardizationofthedotenzymelynkedimmunosorbentassaydotelisaforexperimentalplague
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