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Efficacy of quantitative RT-PCR for detection of the nucleoprotein gene from different porcine rubulavirus strains

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Abstract

Blue-eye disease is an emergent viral swine infection caused by porcine rubulavirus (PoRV). We have developed a qRT-PCR method to detect and quantify expression of the nucleoprotein gene for different PoRV strains. The limit of detection for this assay was 102 copies of synthetic RNA. Viral RNA from PoRV was detectable at a TCID50 of 0.01. Significant differences were observed between viral RNA quantification and virus titration results for nine PoRV strains. For nasal and oral swab samples that were collected from experimentally infected pigs, the qRT-PCR assay was more sensitive (87.1–83.9 %) for the detection of positive samples than methods involving isolation of virus. The implementation of highly sensitive assays that yield results quickly will be of great assistance in the eradication of PoRV from Mexico. We also believe that the newly developed qRT-PCR assay will help reduce the spread of this viral infection to other countries.

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Abbreviations

BED:

Blue-eye disease

CENID-MA:

Centro Nacional de Investigacion Disciplinaria en Microbiologia Animal

IIF:

Indirect immunofluorescence

INIFAP:

Instituto Nacional de Investigaciones Forestales: Agricolas y Pecuarias

LPMV:

La Piedad Michoacan virus

PAC:

Producción Animal Cerdos

PoRV:

Porcine rubulavirus

TCID:

Tissue culture infectious dose

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Acknowledgments

The authors wish to thank Maricela Montalvo-Corral, Monica Resendiz, and Gerardo Santos-López for their aid in the interpretation of our results. The present study was funded by the following projects: PAPIIT-IN224611, CONACYT AC-90024, and CONACYT SSA C02-126709. José F. Rivera-Benitez is the recipient of a grant from the Consejo Nacional de Ciencia y Tecnologia (CONACYT) (ID 207402).

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Facultad de Medicina Veterinaria y Zootecnia, Departamento de Microbiología e Inmunología, Universidad Nacional Autónoma de México, Mexico City, Mexico

    José Francisco Rivera-Benitez & Humberto Ramírez-Mendoza

  2. Departamento de Producción Agrícola y Animal, Universidad Autónoma Metropolitana-Xochimilco, Mexico City, Mexico

    Adelfa del Carmen García-Contreras

  3. Centro de Investigación Biomédica de Oriente, Instituto Mexicano del Seguro Social, Puebla, Mexico

    Julio Reyes-Leyva

  4. Centro de Investigación en Alimentación y Desarrollo, A.C., Sonora, Mexico

    Jesús Hernández

  5. Facultad de Medicina Veterinaria y Zootecnia, Departamento de Medicina y Zootecnia de Cerdos, Universidad Nacional Autónoma de México, Mexico City, Mexico

    José Iván Sánchez-Betancourt

Authors
  1. José Francisco Rivera-Benitez
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  2. Adelfa del Carmen García-Contreras
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  3. Julio Reyes-Leyva
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  4. Jesús Hernández
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  5. José Iván Sánchez-Betancourt
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  6. Humberto Ramírez-Mendoza
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Corresponding author

Correspondence to Humberto Ramírez-Mendoza.

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Rivera-Benitez, J.F., García-Contreras, A.d.C., Reyes-Leyva, J. et al. Efficacy of quantitative RT-PCR for detection of the nucleoprotein gene from different porcine rubulavirus strains. Arch Virol 158, 1849–1856 (2013). https://doi.org/10.1007/s00705-013-1672-0

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  • DOI: https://doi.org/10.1007/s00705-013-1672-0

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