title: Development of a quantitative PCR for rapid and sensitive diagnosis of an intranuclear coccidian parasite in Testudines (TINC), and detection in the critically endangered Arakan forest turtle (Heosemys depressa).
abstract: The intranuclear coccidian parasite of Testudines (TINC) is responsible for significant disease in turtles and tortoises causing high mortality and affecting several threatened species. Diagnostic testing has been limited to relatively labor intensive and expensive pan-coccidial PCR and sequencing techniques. A qPCR assay targeting a specific and conserved region of TINC 18S rRNA was designed. The qPCR reaction was run on samples known to be TINC positive and the results were consistent and analytically specific. The assay was able to detect as little as 10 copies of target DNA in a sample. Testing of soil and invertebrates was negative and did not provide any further insights into life cycles. This assay was used to identify TINC in a novel host species, the critically endangered Arakan forest turtle (Heosemys depressa).
authors.affiliation: Marine Animal Disease Laboratory, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610, USA.
authors.forename: W Alexander
authors.initials: WA
authors.lastname: Alvarez
authors.forename: Paul M
authors.initials: PM
authors.lastname: Gibbons
authors.forename: Sam
authors.initials: S
authors.lastname: Rivera
authors.forename: Linda L
authors.initials: LL
authors.lastname: Archer
authors.forename: April L
authors.initials: AL
authors.lastname: Childress
authors.forename: James F X
authors.initials: JF
authors.lastname: Wellehan
mesh_terms: D000818:Animals; D003048:Coccidiosis / Q000175:diagnosis / Q000662:veterinary*; D056727:Endangered Species; D002052:Myanmar; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D014426:Turtles / Q000469:parasitology*