An improved detection and quantification method for the coral pathogen Vibrio coralliilyticus

DNA- and RNA-based PCR and reverse-transcription real-time PCR assays were developed for diagnostic detection of the vcpA zinc-metalloprotease implicated in the virulence of the coral pathogen Vibrio coralliilyticus. Both PCR methods were highly specific for V. coralliilyticus and failed to amplify...

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Bibliographic Details
Published in:PloS one 2013-12, Vol.8 (12), p.e81800
Main Authors: Wilson, Bryan, Muirhead, Andrew, Bazanella, Monika, Huete-Stauffer, Carla, Vezzulli, Luigi, Bourne, David G
Format: Article
Language:English
Subjects:
Analysis
Animals
Anthozoa - microbiology
Assaying
Bacterial Proteins - genetics
Base Sequence
Bivalvia
Copy number
Deoxyribonucleic acid
Diagnostic systems
Disease
DNA
DNA, Bacterial - genetics
Environmental management
Environmental monitoring
Epidemics
Gene amplification
Genes
Health aspects
Historical account
Metalloproteases - genetics
Metalloproteinase
Methods
Molecular Sequence Data
Mollusca
Pathogenesis
Pathogens
Polymerase chain reaction
Polymerase Chain Reaction - methods
Real time
Ribonucleic acid
RNA
Science
Transcription (Genetics)
Vibrio
Vibrio - genetics
Vibrio - pathogenicity
Vibrio tubiashii
Vibrio vulnificus
Virulence
Virulence (Microbiology)
Waterborne diseases
Zinc
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Summary:DNA- and RNA-based PCR and reverse-transcription real-time PCR assays were developed for diagnostic detection of the vcpA zinc-metalloprotease implicated in the virulence of the coral pathogen Vibrio coralliilyticus. Both PCR methods were highly specific for V. coralliilyticus and failed to amplify strains of closely-related Vibrio species. The assays correctly detected all globally occurring V. coralliilyticus isolates including a newly-described isolate [TAV24] infecting gorgonians in the Mediterranean Sea and highlighted those isolates that had been potentially misidentified, in particular V. tubiashii strains ATCC 19105 and RE22, historically described as important oyster pathogens. The real-time assay is sensitive, detecting 10 gene copies and the relationships between gene copy number and cycle threshold (C T ) were highly linear (R(2)≥ 99.7). The real-time assay was also not affected by interference from non-target DNA. These assays are useful for rapid detection of V. coralliilyticus and monitoring of virulence levels in environmental samples, allowing for implementation of timely management steps to limit and possibly prevent losses due to V. coralliilyticus infection, as well as furthering investigations of factors affecting pathogenesis of this important marine pathogen.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0081800