GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep
Brucellosis is a worldwide zoonosis causing important economic loss and a public health problem. Small ruminants are the preferred hosts of Brucella melitensis and thus the main source of human infections. Effective control of sheep and goat brucellosis has been achieved in several countries through...
| Autores: | , , , , , , , |
|---|---|
| Formato: | artículo |
| Fecha de publicación: | 2018 |
| País: | España |
| Recursos: | Universidad de Navarra |
| Repositorio: | Dadun. Depósito Académico Digital de la Universidad de Navarra |
| Idioma: | inglés |
| OAI Identifier: | oai:dadun.unav.edu:10171/69657 |
| Acesso em linha: | https://hdl.handle.net/10171/69657 |
| Access Level: | acceso abierto |
| Palavra-chave: | Brucella melitensis ELISA-GFP Mini-Tn7-gfp Sheep Vaccines |
| id |
ES_89c480ddd04bc41fb6db9874c2d853fd |
|---|---|
| oai_identifier_str |
oai:dadun.unav.edu:10171/69657 |
| network_acronym_str |
ES |
| network_name_str |
España |
| repository_id_str |
|
| spelling |
GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheepZabalza-Barangua, A. (Ana)|||/items/5d1ba081-59b6-4528-b175-0d64ecf290ebSan-Roman, B. (Beatriz)|||/items/65a686f1-4bd2-47b8-b238-7c064014090aChacon-Diaz, C. (Carlos)|||/items/fd151a1a-c13a-4716-871d-c899310e2bcfde-Miguel, M.J. (María Jesús)|||/items/cfc74851-e6d4-4126-a8f6-71e55b7e72f7Muñoz, P. (Pilar)|||/items/4e98f4c8-d3e4-4fb3-b781-07a0bcb652bcIriarte-Cilveti, M. (Maite)|||/items/5b20b123-3967-4f07-b577-13d13aa57363Blasco, J.M. (José María)|||/items/23b5967d-26c5-43dd-874e-c840176f9aebGrillo, M.J. (María Jesús)|||/items/978ac4b7-4668-4795-9e39-9ba39f15ce07Brucella melitensisELISA-GFPMini-Tn7-gfpSheepVaccinesBrucellosis is a worldwide zoonosis causing important economic loss and a public health problem. Small ruminants are the preferred hosts of Brucella melitensis and thus the main source of human infections. Effective control of sheep and goat brucellosis has been achieved in several countries through vaccination with the live-attenuated B. melitensis Rev1 vaccine. However, Rev1 induces a long-lasting serological response that hinders the differentiation between infected and vaccinated animals. A Rev1::gfp strain expressing constitutively the Green Fluorescent Protein (GFP) was built by stable insertion of a mini-Tn7-gfp in the glmS-recG non-codifying chromosomal region. An associated indirect ELISA-GFP was developed to identify anti-GFP antibodies in vaccinated animals. The resulting Rev1::gfp kept the biological properties of the Rev1 reference strain, including residual virulence and efficacy in mice, and was readily distinguished from Rev1 and other Brucella field strains by direct visualization under ultraviolet illumination, fluorescence microscopy and a multiplex PCR-GFP. The Rev1::gfp strain did not elicit anti-GFP antibodies itself in lambs but when applied in combination with recombinant GFP induced an intense and long-lasting (>9 months) anti-GFP serological response readily detectable by the ELISA-GFP. Overall, our results confirm that Rev1 GFP-tagging can be a suitable alternative for identifying vaccinated sheep in infected contexts.Dadun. Depósito Académico Digital Universidad de Navarra20242024-07-1620182018-01-0120182018-01-01journal articlehttp://purl.org/coar/resource_type/c_6501info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/10171/69657reponame:Dadun. Depósito Académico Digital de la Universidad de Navarrainstname:Universidad de NavarraInglésengopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:dadun.unav.edu:10171/696572026-06-21T12:47:57Z |
| dc.title.none.fl_str_mv |
GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep |
| title |
GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep |
| spellingShingle |
GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep Zabalza-Barangua, A. (Ana)|||/items/5d1ba081-59b6-4528-b175-0d64ecf290eb Brucella melitensis ELISA-GFP Mini-Tn7-gfp Sheep Vaccines |
| title_short |
GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep |
| title_full |
GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep |
| title_fullStr |
GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep |
| title_full_unstemmed |
GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep |
| title_sort |
GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep |
| dc.creator.none.fl_str_mv |
Zabalza-Barangua, A. (Ana)|||/items/5d1ba081-59b6-4528-b175-0d64ecf290eb San-Roman, B. (Beatriz)|||/items/65a686f1-4bd2-47b8-b238-7c064014090a Chacon-Diaz, C. (Carlos)|||/items/fd151a1a-c13a-4716-871d-c899310e2bcf de-Miguel, M.J. (María Jesús)|||/items/cfc74851-e6d4-4126-a8f6-71e55b7e72f7 Muñoz, P. (Pilar)|||/items/4e98f4c8-d3e4-4fb3-b781-07a0bcb652bc Iriarte-Cilveti, M. (Maite)|||/items/5b20b123-3967-4f07-b577-13d13aa57363 Blasco, J.M. (José María)|||/items/23b5967d-26c5-43dd-874e-c840176f9aeb Grillo, M.J. (María Jesús)|||/items/978ac4b7-4668-4795-9e39-9ba39f15ce07 |
| author |
Zabalza-Barangua, A. (Ana)|||/items/5d1ba081-59b6-4528-b175-0d64ecf290eb |
| author_facet |
Zabalza-Barangua, A. (Ana)|||/items/5d1ba081-59b6-4528-b175-0d64ecf290eb San-Roman, B. (Beatriz)|||/items/65a686f1-4bd2-47b8-b238-7c064014090a Chacon-Diaz, C. (Carlos)|||/items/fd151a1a-c13a-4716-871d-c899310e2bcf de-Miguel, M.J. (María Jesús)|||/items/cfc74851-e6d4-4126-a8f6-71e55b7e72f7 Muñoz, P. (Pilar)|||/items/4e98f4c8-d3e4-4fb3-b781-07a0bcb652bc Iriarte-Cilveti, M. (Maite)|||/items/5b20b123-3967-4f07-b577-13d13aa57363 Blasco, J.M. (José María)|||/items/23b5967d-26c5-43dd-874e-c840176f9aeb Grillo, M.J. (María Jesús)|||/items/978ac4b7-4668-4795-9e39-9ba39f15ce07 |
| author_role |
author |
| author2 |
San-Roman, B. (Beatriz)|||/items/65a686f1-4bd2-47b8-b238-7c064014090a Chacon-Diaz, C. (Carlos)|||/items/fd151a1a-c13a-4716-871d-c899310e2bcf de-Miguel, M.J. (María Jesús)|||/items/cfc74851-e6d4-4126-a8f6-71e55b7e72f7 Muñoz, P. (Pilar)|||/items/4e98f4c8-d3e4-4fb3-b781-07a0bcb652bc Iriarte-Cilveti, M. (Maite)|||/items/5b20b123-3967-4f07-b577-13d13aa57363 Blasco, J.M. (José María)|||/items/23b5967d-26c5-43dd-874e-c840176f9aeb Grillo, M.J. (María Jesús)|||/items/978ac4b7-4668-4795-9e39-9ba39f15ce07 |
| author2_role |
author author author author author author author |
| dc.contributor.none.fl_str_mv |
Dadun. Depósito Académico Digital Universidad de Navarra |
| dc.subject.none.fl_str_mv |
Brucella melitensis ELISA-GFP Mini-Tn7-gfp Sheep Vaccines |
| topic |
Brucella melitensis ELISA-GFP Mini-Tn7-gfp Sheep Vaccines |
| description |
Brucellosis is a worldwide zoonosis causing important economic loss and a public health problem. Small ruminants are the preferred hosts of Brucella melitensis and thus the main source of human infections. Effective control of sheep and goat brucellosis has been achieved in several countries through vaccination with the live-attenuated B. melitensis Rev1 vaccine. However, Rev1 induces a long-lasting serological response that hinders the differentiation between infected and vaccinated animals. A Rev1::gfp strain expressing constitutively the Green Fluorescent Protein (GFP) was built by stable insertion of a mini-Tn7-gfp in the glmS-recG non-codifying chromosomal region. An associated indirect ELISA-GFP was developed to identify anti-GFP antibodies in vaccinated animals. The resulting Rev1::gfp kept the biological properties of the Rev1 reference strain, including residual virulence and efficacy in mice, and was readily distinguished from Rev1 and other Brucella field strains by direct visualization under ultraviolet illumination, fluorescence microscopy and a multiplex PCR-GFP. The Rev1::gfp strain did not elicit anti-GFP antibodies itself in lambs but when applied in combination with recombinant GFP induced an intense and long-lasting (>9 months) anti-GFP serological response readily detectable by the ELISA-GFP. Overall, our results confirm that Rev1 GFP-tagging can be a suitable alternative for identifying vaccinated sheep in infected contexts. |
| publishDate |
2018 |
| dc.date.none.fl_str_mv |
2018 2018-01-01 2018 2018-01-01 2024 2024-07-16 |
| dc.type.none.fl_str_mv |
journal article http://purl.org/coar/resource_type/c_6501 |
| dc.type.openaire.fl_str_mv |
info:eu-repo/semantics/article |
| format |
article |
| dc.identifier.none.fl_str_mv |
https://hdl.handle.net/10171/69657 |
| url |
https://hdl.handle.net/10171/69657 |
| dc.language.none.fl_str_mv |
Inglés eng |
| language_invalid_str_mv |
Inglés |
| language |
eng |
| dc.rights.none.fl_str_mv |
open access http://purl.org/coar/access_right/c_abf2 |
| dc.rights.openaire.fl_str_mv |
info:eu-repo/semantics/openAccess |
| rights_invalid_str_mv |
open access http://purl.org/coar/access_right/c_abf2 |
| eu_rights_str_mv |
openAccess |
| dc.format.none.fl_str_mv |
application/pdf |
| dc.source.none.fl_str_mv |
reponame:Dadun. Depósito Académico Digital de la Universidad de Navarra instname:Universidad de Navarra |
| instname_str |
Universidad de Navarra |
| reponame_str |
Dadun. Depósito Académico Digital de la Universidad de Navarra |
| collection |
Dadun. Depósito Académico Digital de la Universidad de Navarra |
| repository.name.fl_str_mv |
|
| repository.mail.fl_str_mv |
|
| _version_ |
1869412651541463040 |
| score |
15,811543 |