Submitted to: American Association of Veterinary Laboratory Diagnosticians
Publication Type: Abstract Only
Publication Acceptance Date: 11/5/2005
Publication Date: 11/5/2005
Citation: Ma, W., Lager, K., Richt, J., Yoon, K. 2005. Development of a multiplex real-time PCR assay for rapid detection and differentiation of PRV [abstract]. American Association of Veterinary Laboratory Diagnosticians 48th Annual Conference. p. 94.
Technical Abstract: Although pseudorabies still exists in many swine producing regions throughout the world, it has recently been eradicated from the U.S. domestic swine herd. A key factor to the successful control and eradication of pseudorabies in the U.S. has been the ability to distinguish animals naturally infected with a wild-type pseudorabies virus (PRV) from vaccinated animals. This was accomplished through the use of gE-deleted modified live virus vaccines and an accompanying differential serologic test (i.e., gE differential ELISA). A critical need for the current PRV surveillance program in the U.S. is the rapid and specific detection of the virus. For this reason, a multiplex real-time PCR using TaqMan chemistry was developed and evaluated for its capability in the detection and differentiation of field and vaccine strains of PRV. Based on sequence information of PRV available in GeneBank and sequences of viruses of all PRV marker vaccines which were commercially available, PCR primers and probes were designed for gB and gE genes for general detection and differentiation, respectively. The PCR assay detected all PRV from diagnostic submissions and vaccine strains in a differential manner. The analytic sensitivity of the assay was approximately 0.1 PFU per reaction. The assay was specific for PRV as no positive results were obtained from testing common swine viral pathogens and other herpesviruses. These observations demonstrated the potential application of the developed multiplex real-time PCR to rapid and specific detection of PRV in domestic and feral swine, as well as in non-porcine species that can be infected with PRV and serve as carriers.