Detection of ovine antibody to Brucella ovis by Indirect Enzyme Immunoassay

Because some batch-to-batch variation in the preparation of rough lipopolysaccharide (RLPS) from Brucella ovis has been experienced, several protocols were tested to establish the most reliable method for detection of antibody in indirect enzyme immunoassay. An early version of the assay gave a perf...

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Autores principales: Nielsen, Klaus, Smith, P., Yu, W. L., Rojas, X., Perez, B., Conde, Sandra Beatriz, Samartino, Luis Ernesto, Robles, Carlos Alejandro
Formato: info:ar-repo/semantics/artículo
Lenguaje:Inglés
Publicado: Taylor & Francis Group 2022
Materias:
Acceso en línea:http://hdl.handle.net/20.500.12123/12414
https://www.tandfonline.com/doi/abs/10.1080/15321810701454789
https://doi.org/10.1080/15321810701454789
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author Nielsen, Klaus
Smith, P.
Yu, W. L.
Rojas, X.
Perez, B.
Conde, Sandra Beatriz
Samartino, Luis Ernesto
Robles, Carlos Alejandro
author_browse Conde, Sandra Beatriz
Nielsen, Klaus
Perez, B.
Robles, Carlos Alejandro
Rojas, X.
Samartino, Luis Ernesto
Smith, P.
Yu, W. L.
author_facet Nielsen, Klaus
Smith, P.
Yu, W. L.
Rojas, X.
Perez, B.
Conde, Sandra Beatriz
Samartino, Luis Ernesto
Robles, Carlos Alejandro
author_sort Nielsen, Klaus
collection INTA Digital
description Because some batch-to-batch variation in the preparation of rough lipopolysaccharide (RLPS) from Brucella ovis has been experienced, several protocols were tested to establish the most reliable method for detection of antibody in indirect enzyme immunoassay. An early version of the assay gave a performance index (PI ¼ sum of optimum percent sensitivity and percent specificity, determined by receiver operator characteristic analysis) of 198.6. This assay used RLPS from B. ovis as the antigen and a monoclonal antibody specific for bovine IgG1 heavy chain-enzyme conjugate for detection. This was not repeatable using other batches of antigen. Newer versions of the assay generally had decreased sensitivity values, giving PIs of 193. Use of a recombinant protein A/G-enzyme conjugate did not improve the PI (PI ¼ 190), giving reduced specificity and higher sensitivity. The final version used B. abortus RB51 RLPS as the antigen and protein A/G-enzyme conjugate for detection, giving a PI of 197. Because of the batch uniformity of the B. abortus RB51 RLPS and the versatility of the protein A/G-enzyme conjugate, the latter version appears to be the most useful for diagnostic serology.
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institution Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina)
language Inglés
publishDate 2022
publishDateRange 2022
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spelling INTA124142022-07-27T11:04:28Z Detection of ovine antibody to Brucella ovis by Indirect Enzyme Immunoassay Nielsen, Klaus Smith, P. Yu, W. L. Rojas, X. Perez, B. Conde, Sandra Beatriz Samartino, Luis Ernesto Robles, Carlos Alejandro Anticuerpos Brucelosis Brucella Brucella Ovis Técnicas Inmunológicas Antibodies Brucellosis Immunological Techniques Inmunoensayo Because some batch-to-batch variation in the preparation of rough lipopolysaccharide (RLPS) from Brucella ovis has been experienced, several protocols were tested to establish the most reliable method for detection of antibody in indirect enzyme immunoassay. An early version of the assay gave a performance index (PI ¼ sum of optimum percent sensitivity and percent specificity, determined by receiver operator characteristic analysis) of 198.6. This assay used RLPS from B. ovis as the antigen and a monoclonal antibody specific for bovine IgG1 heavy chain-enzyme conjugate for detection. This was not repeatable using other batches of antigen. Newer versions of the assay generally had decreased sensitivity values, giving PIs of 193. Use of a recombinant protein A/G-enzyme conjugate did not improve the PI (PI ¼ 190), giving reduced specificity and higher sensitivity. The final version used B. abortus RB51 RLPS as the antigen and protein A/G-enzyme conjugate for detection, giving a PI of 197. Because of the batch uniformity of the B. abortus RB51 RLPS and the versatility of the protein A/G-enzyme conjugate, the latter version appears to be the most useful for diagnostic serology. Estación Experimental Agropecuaria Bariloche Fil: Nielsen, Klaus. Canadian Food Inspection Agency. Ottawa Laboratories; Canadá Fil: Smith, P. Canadian Food Inspection Agency. Ottawa Laboratories; Canadá Fil: Yu, W. L. Canadian Food Inspection Agency. Ottawa Laboratories; Canadá Fil: Rojas, X. Universidad Austral de Chile. Instituto de Microbiología; Chile Fil: Perez, B. Servicio Agrícola y Ganadero; Chile Fil: Conde, Sandra Beatriz. Instituto Nacional de Tecnologia Agropecuaria (INTA). Estación Experimental Agropecuaria Pergamino. Agencia de Extensión Rural Lobos; Argentina Fil: Samartino, Luis Ernesto. Instituto Nacional de Tecnologia Agropecuaria (INTA); Argentina Fil: Robles, Carlos Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Grupo de Sanidad Animal; Argentina 2022-07-27T10:56:32Z 2022-07-27T10:56:32Z 2007 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/12414 https://www.tandfonline.com/doi/abs/10.1080/15321810701454789 1532-1819 1532-4230 https://doi.org/10.1080/15321810701454789 eng info:eu-repo/semantics/restrictedAccess application/pdf Taylor & Francis Group Journal of Immunoassay and Immunochemistry 28 : 243-250 (2007)
spellingShingle Anticuerpos
Brucelosis
Brucella
Brucella Ovis
Técnicas Inmunológicas
Antibodies
Brucellosis
Immunological Techniques
Inmunoensayo
Nielsen, Klaus
Smith, P.
Yu, W. L.
Rojas, X.
Perez, B.
Conde, Sandra Beatriz
Samartino, Luis Ernesto
Robles, Carlos Alejandro
Detection of ovine antibody to Brucella ovis by Indirect Enzyme Immunoassay
title Detection of ovine antibody to Brucella ovis by Indirect Enzyme Immunoassay
title_full Detection of ovine antibody to Brucella ovis by Indirect Enzyme Immunoassay
title_fullStr Detection of ovine antibody to Brucella ovis by Indirect Enzyme Immunoassay
title_full_unstemmed Detection of ovine antibody to Brucella ovis by Indirect Enzyme Immunoassay
title_short Detection of ovine antibody to Brucella ovis by Indirect Enzyme Immunoassay
title_sort detection of ovine antibody to brucella ovis by indirect enzyme immunoassay
topic Anticuerpos
Brucelosis
Brucella
Brucella Ovis
Técnicas Inmunológicas
Antibodies
Brucellosis
Immunological Techniques
Inmunoensayo
url http://hdl.handle.net/20.500.12123/12414
https://www.tandfonline.com/doi/abs/10.1080/15321810701454789
https://doi.org/10.1080/15321810701454789
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