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Novel and highly sensitive sybr® green real-time pcr for poxvirus detection in odontocete cetaceans

Investigación publicada en Journal of Virological Methods

1 de septiembre de 2018

Poxviruses are emerging pathogens in cetaceans, temporarily named `Cetaceanpoxvirus` (CePV, family Poxviridae), classified into two main lineages: CePV-1 in odontocetes and CePV-2 in mysticetes. Only a few studies performed the molecular detection of CePVs, based on DNA-polymerase gene and/or DNA-topoisomerase I gene amplification. Herein we describe a new real-time PCR assay based on SYBR® Green and a new primer set to detect a 150 bp fragment of CePV DNA-polymerase gene, also effective for conventional PCR detection. The novel real-time PCR was able to detect 5 up to 5 × 106 copies per reaction of a cloned positive control. Both novel PCR methods were 1000 to 100,000-fold more sensitive than those previously described in the literature. Samples of characteristic poxvirus skin lesions (`tattoo`) from one Risso`s dolphin (Grampus griseus), two striped dolphins (Stenella coeruleoalba) and two Guiana dolphins (Sotalia guianensis) were all positive to both our novel real time- and conventional PCR methods, even though three of these animals (a Risso`s dolphin, a striped dolphin, and a Guiana dolphin) were previously negative to the conventional PCRs previously available. To our knowledge, this is the first real-time PCR detection method for Cetaceanpoxvirus, a much more sensitive tool for the detection of CePV-1 infections




Sacristan C., Luiz Catao-Dias J., Ewbank AC., Machado EF., Neves E., Santos-Neto EB., Avezedo A., Laison-Brito J., De Castillo PV., Daura-Jorge FG., Simoes-Lopes PC., Carballo M., Garcia-Parraga D., Sanchez-Vizcaino JM. y Esperon F.




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Novel and highly sensitive sybr® green real-time pcr for poxvirus detection in odontocete cetaceans

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Novel and highly sensitive sybr® green real-time pcr for poxvirus detection in odontocete cetaceans


Participantes:

Universidade de Sao PauloLaboratório de Patologia Comparada de Animais Selvagens. Faculdade de Medicina Veterinária e Zootecnia. Universidade de Sao Paulo (USP).

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).

Laboratório de Mamíferos Aquáticos e Bioindicadores (MAQUA). Faculdade De Oceanografia. Universidade do Estado do Rio de Janeiro.

Universidade Federal de Santa CatarinaDepartamento de Engenharia de Pesca. Universidade Federal de Santa Catarina (UFSC).

Universidade Federal de Santa CatarinaDepartamento de Ecologia e Zoologia. Universidade Federal de Santa Catarina (UFSC).

Generalitat ValencianaVeterinary Services. Oceanogràfic. Ciudad de las Artes y las Ciencias. Generalitat Valenciana.

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

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







Journal of Virological Methods
FACTOR YEAR Q
1.746 2018

NLMID: 8005839

PMID: 29890240

ISSN: 0166-0934



TÍTULO: Novel and highly sensitive sybr® green real-time pcr for poxvirus detection in odontocete cetaceans


REVISTA: J. Virol. Methods


NUMERACIÓN: 259:45-49


AÑO: 2018


EDITORIAL: Elsevier/North-Holland Biomedical Press


AUTORES: Sacristan C., Luiz Catao-Dias J., Ewbank AC., Machado EF., Neves E., Santos-Neto EB., Avezedo A., Laison-Brito J., De Castillo PV., Daura-Jorge FG., Simoes-Lopes PC., Carballo M., Garcia-Parraga D., Sanchez-Vizcaino JM. and Esperon F.


PARTICIPANTES VISAVET


José Manuel Sánchez-Vizcaíno Rodríguez

DOI: https://doi.org/10.1016/j.jviromet.2018.06.002


CITA ESTA PUBLICACIÓN:

Sacristan C., Luiz Catao-Dias J., Ewbank AC., Machado EF., Neves E., Santos-Neto EB., Avezedo A., Laison-Brito J., De Castillo PV., Daura-Jorge FG., Simoes-Lopes PC., Carballo M., Garcia-Parraga D., Sanchez-Vizcaino JM. y Esperon F. Novel and highly sensitive sybr® green real-time pcr for poxvirus detection in odontocete cetaceans. Journal of Virological Methods. 259:45-49. 2018. (A). ISSN: 0166-0934. DOI: 10.1016/j.jviromet.2018.06.002


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