Abstract

In order to provide a rapid and sensitive method for detection of the Porcine rubulavirus La Piedad-Michoacan-Mexico Virus (PoRV-LPMV), we have developed a specific real-time reverse transcriptase polymerase chain reaction assay. The detection of PoRV-LPMV, represents a diagnostic challenge due to the viral RNA being present in very small amounts in tissue samples. In this study, a TaqMan (R) real-time PCR assay was designed based on the phosphoprotein gene of PoRV-LPMV, to allow specific amplification and detection of viral RNA in clinical samples. Assay conditions for the primers and probe were optimized using infected PK15 cells and ten-fold serial dilutions of a plasmid containing the whole P-gene. The sensitivity of the developed TaqMan assay was approximately 10 plasmid copies per reaction, and was shown to be 1000 fold better than a conventional nested RT-PCR. The performance of this real-time RT-PCR method enables studies of various aspects of PoRV-LPMV infection. Finally, the assay detects all current known variants of the virus. (C) 2013 Elsevier B.V. All rights reserved.

Keywords

Porcine; Rubulavirus; PoRV-LPMV; Real-time PCR

Published in

Journal of Virological Methods
2013, Volume: 189, number: 1, pages: 1-6
Publisher: ELSEVIER SCIENCE BV

SLU Authors

• Blomström, Anne-Lie

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    

• Berg, Mikael

  • Department of Animal Biosciences, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Pathobiology


Publication identifier

DOI: https://doi.org/10.1016/j.jviromet.2012.12.009

Permanent link to this page (URI)

https://res.slu.se/id/publ/43289