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Simultaneous diagnosis of Cetacean morbillivirus infection in dolphins stranded in the Spanish Mediterranean sea in 2011 using a novel Universal Probe Library (UPL) RT-PCR assay

Investigación publicada en Veterinary Microbiology

19 de enero de 2013

A highly sensitive and specific real-time (rt) RT-PCR assay has been developed for rapid, simultaneous detection of three strains of cetacean morbillivirus (CeMV). In this assay, two PCR primers and a hydrolysis probe from a commercially available Universal Probe Library (UPL) are used to amplify a highly conserved region within the fusion protein gene. RT-PCR is carried out on the same sample using two primer sets in parallel: one set detects the more virulent strains, dolphin morbillivirus (DMV) and porpoise morbillivirus (PMV), and the other set detects the least virulent and least common strain, pilot whale morbillivirus (PWMV). Sensitivity analysis using dilute samples containing purified DMV, PMV and PWMV showed that viral RNA detection limits in this UPL RT-PCR assay were lower than in a conventional RT-PCR assay. Our method gave no amplification signal with field samples positive for viruses related and unrelated to CeMV, such as phocine distemper virus (PDV). The reliability and robustness of the UPL RT-PCR assay were verified using tissue samples previously analyzed by conventional methods, as well as a panel of clinical samples suspected of containing CeMV. Using the UPL RT-PCR assay, we were able to associate DMV with a mass stranding of striped dolphins in the Spanish Mediterranean in 2011 with greater reliability than was possible with a conventional RT-PCR method. These results suggest that this UPL RT-PCR method is more sensitive and specific than the conventional approach, and that it may be an affordable and rapid test for routine diagnosis of three CeMV strains




Rubio-Guerri C., Melero M., Rivera B., Belliere EN., Crespo JL., Garcia-Parraga D., Esperon F. y Sanchez-Vizcaino JM..




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Simultaneous diagnosis of Cetacean morbillivirus infection in dolphins stranded in the Spanish Mediterranean sea in 2011 using a novel Universal Probe Library (UPL) RT-PCR assay

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Simultaneous diagnosis of Cetacean morbillivirus infection in dolphins stranded in the Spanish Mediterranean sea in 2011 using a novel Universal Probe Library (UPL) RT-PCR assay


Participantes:

Universidad ComplutenseServicio de Inmunología Viral y Medicina Preventiva (SUAT). Centro de Vigilancia Sanitaria Veterinaria (VISAVET). Universidad Complutense (UCM).

Ministerio de Ciencia e InnovaciónCentro de Investigación en Sanidad Animal (CISA). Ministerio de Ciencia e Innovación.

Universidad ComplutenseDepartamento de Sanidad Animal. Facultad de Veterinaria. Universidad Complutense (UCM).

Instituto Nacional de Investigación y Tecnología Agraria y AlimentariaCentro de Investigación en Sanidad Animal (CISA). Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA).

Generalitat ValencianaOceanogràfic. Ciudad de las Artes y las Ciencias. Generalitat Valenciana.







Veterinary Microbiology
FACTOR YEAR Q
2.726 2013

NLMID: 7705469

PMID: 23380457

ISSN: 0378-1135



TÍTULO: Simultaneous diagnosis of Cetacean morbillivirus infection in dolphins stranded in the Spanish Mediterranean sea in 2011 using a novel Universal Probe Library (UPL) RT-PCR assay


REVISTA: Vet. Microbiol.


NUMERACIÓN: S0378-1135(13)00040-0


AÑO: 2013


EDITORIAL: Elsevier Science Publishers


AUTORES: Rubio-Guerri C., Melero M., Rivera B., Belliere EN., Crespo JL., Garcia-Parraga D., Esperon F. and Sanchez-Vizcaino JM..


PARTICIPANTES VISAVET


2nd
Mar Melero Asensio
3rd
Belén Rivera Arroyo
Last
José Manuel Sánchez-Vizcaíno Rodríguez

DOI: https://doi.org/10.1016/j.vetmic.2012.12.031


CITA ESTA PUBLICACIÓN:

Rubio-Guerri C., Melero M., Rivera B., Belliere EN., Crespo JL., Garcia-Parraga D., Esperon F. y Sanchez-Vizcaino JM. Simultaneous diagnosis of Cetacean morbillivirus infection in dolphins stranded in the Spanish Mediterranean sea in 2011 using a novel Universal Probe Library (UPL) RT-PCR assay. Veterinary Microbiology. S0378-1135(13)00040-0. 2013. (P). ISSN: 0378-1135. DOI: 10.1016/j.vetmic.2012.12.031


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