Development and validation of a quantitative PCR assay for detection of Sulawesi tortoise adenovirus

Zachary C. Ready, Laura Adamovicz, Maris Daleo, Amber Simmons, Matthew C. Allender

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Article number115033
JournalJournal of Virological Methods
Volume330
DOIs
StatePublished - Dec 2024

Keywords

  • Chelonian
  • Conventional PCR
  • Painted turtle
  • QPCR
  • Sulawesi tortoise adenovirus

ASJC Scopus subject areas

  • Virology

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