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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Virus and Prion Research » Research » Publications at this Location » Publication #379821

Research Project: Intervention Strategies to Control Endemic and New and Emerging Viral Diseases of Swine

Location: Virus and Prion Research

Title: Detection of pseudorabies virus in oral fluid specimen using real-time PCR

item CHENG, TING-YU - Iowa State University
item HARMON, KAREN - Iowa State University
item GAUGER, PHILLIP - Iowa State University
item WANG, CHONG - Iowa State University
item AMBAGALA, ARUNA - Iowa State University
item Buckley, Alexandra
item VAN GEELEN, ALBERT - Animal And Plant Health Inspection Service (APHIS)
item Lager, Kelly
item ZIMMERMAN, JEFFREY - Iowa State University
item GIMENEZ-LIROLA, LUIS - Iowa State University

Submitted to: American Association of Veterinary Laboratory Diagnosticians
Publication Type: Abstract Only
Publication Acceptance Date: 9/1/2020
Publication Date: 10/5/2020
Citation: Cheng, T., Harmon, K., Gauger, P., Wang, C., Ambagala, A., Buckley, A.C., Van Geelen, A., Lager, K.M., Zimmerman, J., Gimenez-Lirola, L. 2020. Detection of pseudorabies virus in oral fluid specimen using real-time PCR [abstract]. American Association of Veterinary Laboratory Diagnosticians. p. not available.

Interpretive Summary:

Technical Abstract: In this study, the detection of PRV DNA in nasal swab ( n = 440) and oral fluid ( n =1,545) samples collected over time from experimentally PRV vaccinated and/or PRV inoculated pigs ( n = 40) was comparatively evaluated by real-time PCR. Serum samples ( n = 440) were tested by PRV gB/gE blocking ELISAs (Pseudorabies Virus gB Antibody Test Kit and Pseudorabies Virus gpI Antibody Test Kit, IDEXX Laboratories, Inc., Westbrook, ME) to monitor PRV status over time. Following exposure to a gE-deleted modified live vaccine (Ingelvac ® Aujeszky MLV, Boehringer Ingelheim, Ridgefield, CT) and/or a wild-type virus (3 CR Ossabaw), PRV gB DNA was detected in oral fluid specimens in a pattern similar to that of nasal swabs. For quantitative analyses, PRV PCR quantification cycle (Cq) results were reexpressed as “efficiency standardized Cqs (ECqs)” as a function of PCR efficiency using plate-specific positive amplification controls. ROC analyses of the PRV gB PCR ECqs results showed a similar performance of the PRV gB PCR for nasal swab and oral fluid specimens (area under the ROC curve = 85% vs 83%) and, based on an ECq cutoff of 0.01 a diagnostic specificity of 100% and diagnostic sensitivities for oral fluid and nasal swab specimens of 53% and 70%, respectively. Thus, the results described herein demonstrated the detection of PRV gB DNA in swine oral fluid and supported the use of this specimen in PRV diagnosis and surveillance.