Análisis de autorías institucional
De Miguel Mj - Autor o Coautor
Muñoz Pm - Autor o Coautor
Blasco Jm - Autor o Coautor
Publicaciones > Artículo

GFP tagging of Brucella melitensis Rev1 allows the identification of vaccinated sheep.

Publicado en:Transboundary And Emerging Diseases. 66 (1): 505-516 - 2019-01-01 66(1), doi: 10.1111/tbed.13053

De Miguel Mj; Muñoz Pm; Blasco Jm;


2010CR0005 AGL2011?30453?C04?01, RTC?2015? 3618?1, AGL2014?58795?C04 Universidad Pública de Navarra - Financiador
Centro de Investigación en Enfermedades Tropicales, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica. - Autor o Coautor
Centro de Investigación y Tecnología Agroalimentaria (CITA), Instituto Agroalimentario de Aragón (IA2), Gobierno de Aragón, Zaragoza, Spain. - Autor o Coautor
Centro de Investigación y Tecnología Agroalimentaria de Aragón - Entidad de origen
Instituto de Agrobiotecnología (IdAB, CSIC-Gobierno de Navarra), Mutilva, Navarra, Spain. - Autor o Coautor
Instituto de Salud Tropical - Dpto. de Microbiología y Parasitología, Universidad de Navarra, Pamplona, Spain. - Autor o Coautor
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Resúmen: 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.© 2018 Blackwell Verlag GmbH.

Palabras clave: brucella melitensis ; Brucella melitensis; Elisa-gfp; Mini-tn7-gfp; Sheep; Vaccines

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