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 throu...
| Main Authors: | , , , , , , , |
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| Format: | info:eu-repo/semantics/article |
| Language: | eng |
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Wiley
2022
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| Online Access: | https://hdl.handle.net/10171/63630 |
| _version_ | 1793400397700792320 |
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| author | Zabalza-Barangua, A. (Ana) San-Roman, B. (Beatriz) Chacon-Diaz, C. (Carlos) Miguel, M.J. (María Jesús) de Muñoz, P. (Pilar) Iriarte, M. (Maite) Blasco, J.M. (J. M.) Grillo, M.J. (María Jesús) |
| author_facet | Zabalza-Barangua, A. (Ana) San-Roman, B. (Beatriz) Chacon-Diaz, C. (Carlos) Miguel, M.J. (María Jesús) de Muñoz, P. (Pilar) Iriarte, M. (Maite) Blasco, J.M. (J. M.) Grillo, M.J. (María Jesús) |
| author_sort | Zabalza-Barangua, A. (Ana) |
| collection | DSpace |
| 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 | info:eu-repo/semantics/article |
| id | oai:dadun.unav.edu:10171-63630 |
| institution | Universidad de Navarra |
| language | eng |
| publishDate | 2022 |
| publisher | Wiley |
| record_format | dspace |
| spelling | oai:dadun.unav.edu:10171-636302022-06-10T01:05:15Z GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep Zabalza-Barangua, A. (Ana) San-Roman, B. (Beatriz) Chacon-Diaz, C. (Carlos) Miguel, M.J. (María Jesús) de Muñoz, P. (Pilar) Iriarte, M. (Maite) Blasco, J.M. (J. M.) Grillo, M.J. (María Jesús) Brucella melitensis ELISA-GFP Mini-Tn7-gfp Sheep Vaccines 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. 2022-06-09T12:05:15Z 2022-06-09T12:05:15Z 2019 info:eu-repo/semantics/article https://hdl.handle.net/10171/63630 eng info:eu-repo/semantics/openAccess application/pdf Wiley |
| spellingShingle | Brucella melitensis ELISA-GFP Mini-Tn7-gfp Sheep Vaccines Zabalza-Barangua, A. (Ana) San-Roman, B. (Beatriz) Chacon-Diaz, C. (Carlos) Miguel, M.J. (María Jesús) de Muñoz, P. (Pilar) Iriarte, M. (Maite) Blasco, J.M. (J. M.) Grillo, M.J. (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 | Brucella melitensis ELISA-GFP Mini-Tn7-gfp Sheep Vaccines |
| url | https://hdl.handle.net/10171/63630 |
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