Development of a Quantitative PCR Assay for a Novel Adenovirus in the Box Turtle (Terrapene carolina)
American Association of Zoo Veterinarians Conference 2014
Stella A. Blum1; Terry Norton2, DVM, DACZM; Sharon Deem3, DVM, PhD, DACZM; Natalie Hall4, DVM, DACZM; Greg Fleming4, DVM, DACZM; James F.X. Wellehan Jr.5, DVM, MS, PhD, DACZM
1College of Veterinary Medicine, University of Florida, Gainesville, FL, USA; 2Georgia Sea Turtle Center, Jekyll Island, GA, USA; 3St. Louis Zoo Institute for Conservation Medicine, St. Louis, MO, USA; 4Disney’s Animal Programs, Orlando, FL, USA; 5Zoological Medicine Service, Department of Small Animal Clinical Sciences, University of Florida, Gainesville, FL, USA
Abstract
Historically, a number of mortality events have been observed in Eastern box turtles with iridoviruses and other unknown causes. Novel adenoviruses have recently been detected by consensus PCR in several chelonian species, including Eastern box turtles (Terrapene carolina). This species is highly vulnerable to population declines from the loss of a relatively small number of adults. Diagnostic testing has been limited to relatively labor-intensive consensus PCR techniques. A qPCR assay targeting a specific and conserved region of the DNA polymerase gene of the novel Box turtle adenovirus 1 was designed to allow for rapid and sensitive diagnosis. The qPCR reaction was run on several samples determined by consensus PCR to be box turtle adenovirus 1 positive, and was able to detect as little as 10 counts of DNA in a sample. The results were consistent and analytically specific to this virus. The assay was used to screen wild and captive populations of box turtles for the virus.