GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep.
Animals
Brucella Vaccine
/ administration & dosage
Brucella melitensis
/ immunology
Brucellosis
/ immunology
Enzyme-Linked Immunosorbent Assay
/ veterinary
Female
Green Fluorescent Proteins
/ immunology
Immunoglobulin G
/ blood
Luminescent Agents
Male
Mice
Mice, Inbred BALB C
Microscopy, Fluorescence
/ veterinary
Sheep
Sheep Diseases
/ diagnosis
Vaccination
/ veterinary
Vaccines, Attenuated
/ immunology
Brucella melitensis
ELISA-GFP
mini-Tn7-gfp
sheep
vaccines
Journal
Transboundary and emerging diseases
ISSN: 1865-1682
Titre abrégé: Transbound Emerg Dis
Pays: Germany
ID NLM: 101319538
Informations de publication
Date de publication:
Jan 2019
Jan 2019
Historique:
received:
02
07
2018
revised:
17
10
2018
accepted:
22
10
2018
pubmed:
31
10
2018
medline:
5
3
2019
entrez:
31
10
2018
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 30375177
doi: 10.1111/tbed.13053
pmc: PMC7379934
doi:
Substances chimiques
Brucella Vaccine
0
Immunoglobulin G
0
Luminescent Agents
0
Vaccines, Attenuated
0
Green Fluorescent Proteins
147336-22-9
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
505-516Subventions
Organisme : Secretaría de Estado de Investigación, Desarrollo e Innovación
ID : AGL2011-30453-C04-01
Organisme : Secretaría de Estado de Investigación, Desarrollo e Innovación
ID : AGL2014-58795-C04
Organisme : Secretaría de Estado de Investigación, Desarrollo e Innovación
ID : RTC-2015-3618-1
Organisme : CSIC-CRUSA Foundation bilateral project
ID : 2010CR0005
Informations de copyright
© 2018 Blackwell Verlag GmbH.
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