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dc.contributor.authorDeFaveri, Jacquelin
dc.contributor.authorSmolowitz, Roxanna M.
dc.contributor.authorRoberts, Steven B.
dc.date.accessioned2010-04-14T14:19:18Z
dc.date.available2010-04-14T14:19:18Z
dc.date.issued2009-08
dc.identifier.citationJournal of Shellfish Research 28 (2009): 459-464en_US
dc.identifier.urihttp://hdl.handle.net/1912/3241
dc.descriptionAuthor Posting. © National Shellfisheries Association, 2009. This article is posted here by permission of National Shellfisheries Association for personal use, not for redistribution. The definitive version was published in Journal of Shellfish Research 28 (2009): 459-464, doi:10.2983/035.028.0306.en_US
dc.description.abstractPerkinus marinus causes a devastating disease, known as Dermo, in the Eastern oyster Crassostrea virginica. Routine detection of the disease is traditionally accomplished by the use of the Ray/Makin assay, using Fluid Thioglycollate Medium (RFTM). A simple real-time quantitative PCR assay was developed as a diagnostic tool to detect and quantify P. marinus, to complement and serve as an alternate to the RFTM method. Using a dual-labeled probe approach, a sensitive assay was designed to accurately detect a range of one to several thousand P. marinus organisms present in oyster tissues. A simple extraction method was used to increase throughput of the assay. Cultured P. marinus cells were quantified prior to DNA extraction, generating a standard curve and allowing cell counts to be derived from PCR cycle threshold values. Direct comparison of the RFTM and real-time PCR methods was accomplished by using tissue samples from the same oyster for both tests. Plotting cycle threshold values against the known Mackin index value generated a standard curve with a coefficient of regression of 0.9. Our results indicate that correlations could be made between this molecular based approach and traditional methods, allowing results generated from the PCR assay to be easily translated into the understood Mackin scale.en_US
dc.format.mimetypeapplication/pdf
dc.language.isoen_USen_US
dc.publisherNational Shellfisheries Associationen_US
dc.relation.urihttps://doi.org/10.2983/035.028.0306
dc.subjectDermoen_US
dc.subjectPerkinsus marinusen_US
dc.subjectReal-time PCRen_US
dc.subjectRFTMen_US
dc.subjectEastern oysteren_US
dc.subjectCrassostrea virginicaen_US
dc.titleDevelopment and validation of a real-time quantitative PCR assay for the detection and quantification of Perkinsus marinus in the Eastern oyster, Crassostrea virginicaen_US
dc.typeArticleen_US
dc.identifier.doi10.2983/035.028.0306


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