Development of a real time quantitative PCR assay for the hard clam pathogen Quahog Parasite Unknown (QPX).

Development of a real time quantitative PCR assay for the hard clam pathogen Quahog Parasite Unknown (QPX). Research Abstract Details 

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Development of a real time quantitative PCR assay for the hard clam pathogen Quahog Parasite Unknown (QPX). Abstract Text:

M Maille Lyons,Roxanna Smolowitz,Christopher F Dungan,Steven B Roberts,

Quahog Parasite Unknown (QPX) is a thraustochytrid pathogen responsible for catastrophic mortalities of the northern quahog (hard clam) Mercenaria mercenaria. A real-time quantitative polymerase chain reaction (qPCR) assay was developed to assist research efforts on QPX ecology and pathology. Sensitivity of the assay was evaluated with serial dilutions of QPX-cultured cells to determine the lowest concentration of DNA that remained detectable in both the presence and absence of extraneous environmental substances. QPX cells were quantified before DNA extraction to calibrate standard curves to cell counts. Based on our results, the qPCR assay is able to quantify QPX within the range of 1 to several thousand organisms per reaction. Specificity of the assay was assessed by testing 29 thraustochytrid-like protists isolated from suspension-feeding bivalves from China, Oregon, Maryland, and Virginia. Application of the assay was demonstrated with positive qPCR results from naturally contaminated environmental samples including marine aggregates (i.e. marine snow), clam pseudofeces, and inflammatory nodules from infected clams. This quantitative assay for QPX will provide a valuable tool for characterizing QPX parasite abundances in coastal environments and for improving clam disease diagnostics.

Development of a real time quantitative PCR assay for the hard clam pathogen Quahog Parasite Unknown (QPX). Publishing Authors By Initials

MM Lyons,R Smolowitz,CF Dungan,SB Roberts,

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Development of a real time quantitative PCR assay for the hard clam pathogen Quahog Parasite Unknown (QPX). Journal Published:

PUBLICATION TYPE: Research Support, U.S. Gov't,

Journal: Diseases of aquatic organisms

VOLUME: 72

Page Numbers: 45-52

Journal Abbreviation: Dis. Aquat. Org.

ISSN: 0177-5103

DAY: 14

MONTH: Sep

YEAR: 2006

Development of a real time quantitative PCR assay for the hard clam pathogen Quahog Parasite Unknown (QPX). Information

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LANGUAGE: eng

NlmUniqueID: 8807037

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Grant and Affiliation Information for Development of a real time quantitative PCR assay for the hard clam pathogen Quahog Parasite Unknown (QPX).

AFFILIATION: Department of Marine Sciences, University of Connecticut, 1080 Shennecossett Road Groton, Connecticut 06340, USA.

Country: Germany

AGENCY: United States PHS

GRANT: 0429018

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MEDLINETA: Dis Aquat Organ

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