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

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 10 2 copies of synthetic RNA. Viral RNA fro...

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Bibliographic Details
Published in:Archives of virology 2013-09, Vol.158 (9), p.1849-1856
Main Authors: Rivera-Benitez, José Francisco, García-Contreras, Adelfa del Carmen, Reyes-Leyva, Julio, Hernández, Jesús, Sánchez-Betancourt, José Iván, Ramírez-Mendoza, Humberto
Format: Article
Language:English
Subjects:
Animals
Biomedical and Life Sciences
Biomedicine
Cell culture
Eye diseases
Genotype
Hogs
Infection
Infections
Infectious Diseases
Medical Microbiology
Methods
Mexico
Neurological disorders
Nucleoproteins - genetics
Nucleoproteins - metabolism
Original Article
Porcine rubulavirus
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction - methods
RNA, Viral - analysis
RNA, Viral - genetics
Rubulavirus - classification
Rubulavirus - genetics
Rubulavirus - isolation & purification
Rubulavirus Infections - veterinary
Rubulavirus Infections - virology
Sensitivity and Specificity
Serology
Swine
Swine Diseases - virology
Viral infections
Viral Proteins - genetics
Viral Proteins - metabolism
Virology
Viruses
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Summary: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 10 2 copies of synthetic RNA. Viral RNA from PoRV was detectable at a TCID 50 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.
ISSN:0304-8608
1432-8798
DOI:10.1007/s00705-013-1672-0