Development and validation of TaqMan quantitative PCR for detection of frog virus 3-like virus in eastern box turtles (Terrapene carolina carolina)
► 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. Ranavirus has caused disease epidemics and mass mortality events globally in fre...
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Published in | Journal of virological methods Vol. 188; no. 1-2; pp. 121 - 125 |
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Main Authors | , , |
Format | Journal Article |
Language | English |
Published |
Netherlands
Elsevier B.V
01.03.2013
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Subjects | |
Online Access | Get full text |
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Summary: | ► 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.
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. |
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Bibliography: | http://dx.doi.org/10.1016/j.jviromet.2012.12.012 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0166-0934 1879-0984 |
DOI: | 10.1016/j.jviromet.2012.12.012 |