Literaturdatenbank
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Allender, M. C., Bunick, D., & Mitchell, M. A. (2013). Development and validation of taqman quantitative pcr for detection of frog virus 3-like virus in eastern box turtles (terrapene carolina carolina). Journal of Virological Methods, (in press, uncorrected proof).
Added by: Admin (06 Jan 2014 18:22:35 UTC) |
| Resource type: Journal Article BibTeX citation key: Allender2013 View all bibliographic details  |
Categories: General Keywords: Emydidae, Schildkröten - turtles + tortoises, Terrapene carolina, Veterinärmedizin - veterinary medicine, Viren - viruses Creators: Allender, Bunick, Mitchell Collection: Journal of Virological Methods |
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| Abstract |
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Terrapene carolina 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. Highlights ► qPCR is 1000 more sensitive than conventional PCR. ► Assay has similar efficiency in plasmid, cell lysates, and dilutions of whole turtle blood. ► Assay targets a conservative portion of the major capsid protein gene.
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