Bacterial expressed coat protein: development of a single antiserum for routine detection of Citrus tristeza virus

Detalhes bibliográficos
Autor(a) principal: Sequeira, Z.
Data de Publicação: 2002
Outros Autores: Nolasco, Gustavo
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10400.1/6244
Resumo: Citrus tristeza virus (CTV) causes one of the most important citrus diseases world-wide and has recently been detected in Portugal. Early diagnosis based on immunoenzymatic techniques requires significant amounts of reagents. This research describes the isolation of the coat protein gene of CTV, its expression in Escherichia coli as a fusion protein containing an N-terminal (His)6 region and its use to raise rabbit polyclonal antibodies. These antibodies could be used at dilutions higher than 1/80,000 at the detection stage in indirect ELISA tests and were also efficient for trapping the virus in ELISA and Immunocapture RT-PCR. These characteristics allowed the production of diagnostic kits based solely on this source of antibodies. A detection spectrum and sensitivity similar to that of a commercial polyclonal antibody kit was achieved.
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spelling Bacterial expressed coat protein: development of a single antiserum for routine detection of Citrus tristeza virusCitrus tristeza virus (CTV) causes one of the most important citrus diseases world-wide and has recently been detected in Portugal. Early diagnosis based on immunoenzymatic techniques requires significant amounts of reagents. This research describes the isolation of the coat protein gene of CTV, its expression in Escherichia coli as a fusion protein containing an N-terminal (His)6 region and its use to raise rabbit polyclonal antibodies. These antibodies could be used at dilutions higher than 1/80,000 at the detection stage in indirect ELISA tests and were also efficient for trapping the virus in ELISA and Immunocapture RT-PCR. These characteristics allowed the production of diagnostic kits based solely on this source of antibodies. A detection spectrum and sensitivity similar to that of a commercial polyclonal antibody kit was achieved.SapientiaSequeira, Z.Nolasco, Gustavo2015-06-15T14:13:47Z20022002-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.1/6244eng0031-9465AUT: GNO00324;info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-24T10:17:27Zoai:sapientia.ualg.pt:10400.1/6244Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:59:03.118020Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Bacterial expressed coat protein: development of a single antiserum for routine detection of Citrus tristeza virus
title Bacterial expressed coat protein: development of a single antiserum for routine detection of Citrus tristeza virus
spellingShingle Bacterial expressed coat protein: development of a single antiserum for routine detection of Citrus tristeza virus
Sequeira, Z.
title_short Bacterial expressed coat protein: development of a single antiserum for routine detection of Citrus tristeza virus
title_full Bacterial expressed coat protein: development of a single antiserum for routine detection of Citrus tristeza virus
title_fullStr Bacterial expressed coat protein: development of a single antiserum for routine detection of Citrus tristeza virus
title_full_unstemmed Bacterial expressed coat protein: development of a single antiserum for routine detection of Citrus tristeza virus
title_sort Bacterial expressed coat protein: development of a single antiserum for routine detection of Citrus tristeza virus
author Sequeira, Z.
author_facet Sequeira, Z.
Nolasco, Gustavo
author_role author
author2 Nolasco, Gustavo
author2_role author
dc.contributor.none.fl_str_mv Sapientia
dc.contributor.author.fl_str_mv Sequeira, Z.
Nolasco, Gustavo
description Citrus tristeza virus (CTV) causes one of the most important citrus diseases world-wide and has recently been detected in Portugal. Early diagnosis based on immunoenzymatic techniques requires significant amounts of reagents. This research describes the isolation of the coat protein gene of CTV, its expression in Escherichia coli as a fusion protein containing an N-terminal (His)6 region and its use to raise rabbit polyclonal antibodies. These antibodies could be used at dilutions higher than 1/80,000 at the detection stage in indirect ELISA tests and were also efficient for trapping the virus in ELISA and Immunocapture RT-PCR. These characteristics allowed the production of diagnostic kits based solely on this source of antibodies. A detection spectrum and sensitivity similar to that of a commercial polyclonal antibody kit was achieved.
publishDate 2002
dc.date.none.fl_str_mv 2002
2002-01-01T00:00:00Z
2015-06-15T14:13:47Z
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