A triplex real-time PCR for differential detection of classical, variant and Bartha-K61 vaccine strains of pseudorabies virus

A triplex real-time PCR for differential detection of classical, variant and Bartha-K61 vaccine... Pseudorabies (PR), also known as Aujeszky’s disease, is an economically important infectious disease of pigs and other animals caused by pseudorabies virus (PRV). Since late 2011, increasing numbers of PR outbreaks have been reported on many Bartha-K61-vaccinated pig farms in China, and emerging PRV variants that differ from classical PRV strains genetically and antigenically have been confirmed to be responsible for the outbreaks. Accordingly, there is a need to differentiate diverse PRV strains co-circulating in the field. Here, we developed and evaluated a triplex real-time PCR for differential detection of wild-type PRV (classical and variant strains) and gE/gI gene-deleted vaccine strains based on three differently labeled TaqMan probes. The detection limits of the assay were 0.5 TCID50 for classical strains, 0.2 TCID50 for variant strains and 0.05 TCID50 for vaccine strains. The sensitivity was also determined to be 50, 50 and 5 copies for the TJ, SC and Bartha-K61 strain, respectively. The assay did not show cross-reactivity with several common porcine viruses. Reproducibility tests showed that the inter- and intra-assay coefficients of variation were less than 3 %. When testing a total of 234 clinical swine samples, the agreement between the triplex real-time PCR and virus isolation was 100 % (234/234) for classical strains, 99.5 % (233/234) for variant strains, and 100 % (234/234) for the Bartha-K61 vaccine strain. The results demonstrate that this method is sensitive and specific and will be useful for rapid detection and differentiation of diverse PRV strains. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

A triplex real-time PCR for differential detection of classical, variant and Bartha-K61 vaccine strains of pseudorabies virus

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Publisher
Springer Vienna
Copyright
Copyright © 2016 by Springer-Verlag Wien
Subject
Biomedicine; Virology; Medical Microbiology; Infectious Diseases
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s00705-016-2925-5
Publisher site
See Article on Publisher Site

Abstract

Pseudorabies (PR), also known as Aujeszky’s disease, is an economically important infectious disease of pigs and other animals caused by pseudorabies virus (PRV). Since late 2011, increasing numbers of PR outbreaks have been reported on many Bartha-K61-vaccinated pig farms in China, and emerging PRV variants that differ from classical PRV strains genetically and antigenically have been confirmed to be responsible for the outbreaks. Accordingly, there is a need to differentiate diverse PRV strains co-circulating in the field. Here, we developed and evaluated a triplex real-time PCR for differential detection of wild-type PRV (classical and variant strains) and gE/gI gene-deleted vaccine strains based on three differently labeled TaqMan probes. The detection limits of the assay were 0.5 TCID50 for classical strains, 0.2 TCID50 for variant strains and 0.05 TCID50 for vaccine strains. The sensitivity was also determined to be 50, 50 and 5 copies for the TJ, SC and Bartha-K61 strain, respectively. The assay did not show cross-reactivity with several common porcine viruses. Reproducibility tests showed that the inter- and intra-assay coefficients of variation were less than 3 %. When testing a total of 234 clinical swine samples, the agreement between the triplex real-time PCR and virus isolation was 100 % (234/234) for classical strains, 99.5 % (233/234) for variant strains, and 100 % (234/234) for the Bartha-K61 vaccine strain. The results demonstrate that this method is sensitive and specific and will be useful for rapid detection and differentiation of diverse PRV strains.

Journal

Archives of VirologySpringer Journals

Published: Jun 17, 2016

References

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