Bacterial expressed coat protein: development of a single antiserum for routine detection of Citrus tristeza virus
Autor(a) principal: | |
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Data de Publicação: | 2002 |
Outros Autores: | |
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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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 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.1/6244 |
url |
http://hdl.handle.net/10400.1/6244 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
0031-9465 AUT: GNO00324; |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799133210591887360 |