Development and validation of TaqMan quantitative PCR for detection of frog virus 3-like virus in eastern box turtles (Terrapene carolina carolina)

Matthew C. Allender, David Bunick, Mark A. Mitchell

Research output: Contribution to journalArticle

Abstract

Ranavirus has caused disease epidemics and mass mortality events globally in free-ranging fish, amphibian, and reptile populations. Viral isolation and conventional PCR are the most common methods for diagnosis. In this study, a quantitative real-time PCR (qPCR) assay was developed using a TaqMan probe-based assay targeting a highly conserved region of the major capsid protein of frog virus 3-like virus (FV3-like) (Family Iridoviridae, genera Ranavirus). Standard curves were generated from a viral DNA segment cloned within a plasmid. The assay detected viral DNA 1000 times lower than conventional PCR. Thirty-one clinical samples (whole blood and oral swabs) from box turtles were tested using these assays and the prevalence of the virus determined. Quantitative PCR allows for a superior, rapid, sensitive, and quantitative method for detecting FV3-like virus in box turtles, and this assay will be useful for early detection and disease monitoring.

Original languageEnglish (US)
Pages (from-to)121-125
Number of pages5
JournalJournal of Virological Methods
Volume188
Issue number1-2
DOIs
StatePublished - Mar 1 2013

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Keywords

  • Epidemiology
  • Molecular
  • Ranavirus
  • Reptile

ASJC Scopus subject areas

  • Virology

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