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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 principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7379934/ https://www.ncbi.nlm.nih.gov/pubmed/30375177 http://dx.doi.org/10.1111/tbed.13053 |
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author | Zabalza‐Baranguá, Ana San‐Román, Beatriz Chacón‐Díaz, Carlos de Miguel, María‐Jesús Muñoz, Pilar‐María Iriarte, Maite Blasco, José‐María Grilló, María‐Jesús |
author_facet | Zabalza‐Baranguá, Ana San‐Román, Beatriz Chacón‐Díaz, Carlos de Miguel, María‐Jesús Muñoz, Pilar‐María Iriarte, Maite Blasco, José‐María Grilló, María‐Jesús |
author_sort | Zabalza‐Baranguá, Ana |
collection | PubMed |
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. |
format | Online Article Text |
id | pubmed-7379934 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-73799342020-07-27 GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep Zabalza‐Baranguá, Ana San‐Román, Beatriz Chacón‐Díaz, Carlos de Miguel, María‐Jesús Muñoz, Pilar‐María Iriarte, Maite Blasco, José‐María Grilló, María‐Jesús Transbound Emerg Dis Original Articles 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. John Wiley and Sons Inc. 2018-11-26 2019-01 /pmc/articles/PMC7379934/ /pubmed/30375177 http://dx.doi.org/10.1111/tbed.13053 Text en © 2018 The Authors. Transboundary and Emerging Diseases published by Blackwell Verlag GmbH This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Articles Zabalza‐Baranguá, Ana San‐Román, Beatriz Chacón‐Díaz, Carlos de Miguel, María‐Jesús Muñoz, Pilar‐María Iriarte, Maite Blasco, José‐María Grilló, María‐Jesús 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 |
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_short |
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 |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7379934/ https://www.ncbi.nlm.nih.gov/pubmed/30375177 http://dx.doi.org/10.1111/tbed.13053 |
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