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Updated: Jun 12, 2025
Author Spotlight: Addressing Regulatory Gaps in Molecular Studies by Quantifying Viral Vectors in Complex Matrices
Published on: July 14, 2023
Development and validation of a quantitative PCR assay for detection of Sulawesi tortoise adenovirus
Zachary C Ready1, Laura Adamovicz2, Maris Daleo2
1Illinois Zoological and Aquatic Animal Residency, Shedd Aquarium, Chicago, IL 60605, USA; University of Illinois College of Veterinary Medicine, Urbana, IL 61802, USA; Chicago Zoological Society, Brookfield Zoo, Brookfield, IL 60513, USA; Wildlife Epidemiology Laboratory, University of Illinois, Urbana, IL 61802, USA.
Abstract:
In 2007, a mortality event involving over 100 Sulawesi tortoises (Indotestudo forsteni), two Impressed tortoises (Manouria impress) and a critically endangered Burmese star tortoise (Geochelone platynota) was attributed to Sulawesi tortoise adenovirus (STADV; genus Siadenovirus). We developed a TaqMan quantitative PCR assay targeting the DNA polymerase gene of STADV for use in clinical diagnosis and epidemiologic surveillance. This assay failed to amplify five closely-related chelonian adenoviruses, indicating high analytical specificity. The assay performed with high efficiency (slope = -3.337; R2 = 0.999) and high inter- and intra-assay repeatability (coefficient of variation <1.36 % at all standard curve dilutions). Dynamic range included 1.00 × 107 to 1.00 × 101 target copies per reaction and limit of detection was 101 target copies per reaction, though 100 target copies per reaction were intermittently detected. This qPCR assay provides a valuable diagnostic tool for characterization of STADV epidemiology, including potential identification of the North American reservoir host.
