PMID- 17465139
OWN - NLM
STAT- MEDLINE
DCOM- 20070810
LR  - 20131121
IS  - 0034-7744 (Print)
IS  - 0034-7744 (Linking)
VI  - 53 Suppl 1
DP  - 2005 May
TI  - Development and field application of a molecular probe for the primary pathogen 
      of the coral disease white plague type II.
PG  - 1-10
AB  - One of the current problems in the field of coral disease research is that of 
      tracking coral pathogens in the natural environment. A promising method to do 
      this is by use of pathogen-specific molecular probes. However, this approach has 
      been little used to date. We constructed, and validated in the laboratory, a 
      fluorochrome-labeled molecular probe specific to Aurantimonas coralicida, the 
      bacterial pathogen of the Caribbean coral disease white plague type II (WPIl). We 
      then used the probe to test field samples of diseased coral tissue for the 
      presence of this pathogen. Probe design was based on a unique subset (25 
      nucleotides) of the complete l6S rRNA gene sequence derived from a pure culture 
      of the pathogen. The pathogen-specific probe was labeled with the fluorochrome 
      GreenStar* FITC (fluorescein isothiocyanate, GeneDetect Ltd, New Zealand). As a 
      control, we used the universal eubacterial probe EUB 338, labeled with a 
      different fluorochrome (TRITC, tetra-methylrhodamine isothiocyanate). Both probes 
      were applied to laboratory samples of pure cultures of bacteria, and field 
      samples collected from the surface of the disease line of corals exhibiting signs 
      of white plague (types I and II), healthy controls, and corals with an 
      uncharacterized disease ("patchy necrosis"). All samples were analyzed using 
      fluorescence in situ hybridization (FISH). We have determined that the probe is 
      specific to our laboratory culture of the coral pathogen, and does not react with 
      other bacterial species (the eubacterial probe does). The WPII pathogen was 
      detected in association with diseased coral samples collected from coral colonies 
      on reefs of the Bahamas (n= 9 samples) exhibiting signs of both WPI and WPII. 
      Diseased (and healthy) tissue samples (n- 4) from corals exhibiting signs of 
      "patchy necrosis" were also assayed. In this case the results were negative, 
      indicating that the same pathogen is not involved in the two diseases. 
      Incorporation and use of pathogen-specific probes can significantly expand our 
      knowledge of the etiology of coral diseases.
FAU - Richardson, Laurie L
AU  - Richardson LL
AD  - Department of Biological Sciences, Florida International University, Miami, 
      Florida 33199, USA. Laurie.Richardson@fiu.edu
FAU - Mills, DeEtta K
AU  - Mills DK
FAU - Remily, Elizabeth R
AU  - Remily ER
FAU - Voss, Joshua D
AU  - Voss JD
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - Costa Rica
TA  - Rev Biol Trop
JT  - Revista de biologia tropical
JID - 0404267
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Molecular Probes)
RN  - 0 (RNA, Ribosomal, 16S)
RN  - I223NX31W9 (Fluorescein-5-isothiocyanate)
SB  - IM
MH  - Animals
MH  - Anthozoa/chemistry/genetics/*microbiology
MH  - Colony Count, Microbial
MH  - Fluorescein-5-isothiocyanate/*analysis
MH  - Fluorescent Dyes/*analysis
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Molecular Probe Techniques/*instrumentation
MH  - Molecular Probes/genetics
MH  - Necrosis/genetics/pathology
MH  - RNA, Ribosomal, 16S/genetics
MH  - Rhizobiaceae/*isolation & purification
MH  - Sensitivity and Specificity
EDAT- 2007/05/01 09:00
MHDA- 2007/08/11 09:00
CRDT- 2007/05/01 09:00
PHST- 2007/05/01 09:00 [pubmed]
PHST- 2007/08/11 09:00 [medline]
PHST- 2007/05/01 09:00 [entrez]
PST - ppublish
SO  - Rev Biol Trop. 2005 May;53 Suppl 1:1-10.