PMID- 22443397
OWN - NLM
STAT- MEDLINE
DCOM- 20121126
LR  - 20181201
IS  - 1365-2672 (Electronic)
IS  - 1364-5072 (Linking)
VI  - 112
IP  - 6
DP  - 2012 Jun
TI  - Development of a new CARD-FISH protocol for quantification of Legionella 
      pneumophila and its application in two hospital cooling towers.
PG  - 1244-56
LID - 10.1111/j.1365-2672.2012.05289.x [doi]
AB  - AIMS: Open cooling towers are frequent sources of infections with Legionella 
      pneumophila. The gold standard for the detection of Leg. pneumophila is based on 
      cultivation lasting up to 10 days and detecting only culturable cells. 
      Alternative fluorescence in situ hybridization (FISH) protocols have been 
      proposed, but they result in faint fluorescence signals and lack specificity 
      because of cross-hybridization with other Legionella species. Our aim was thus to 
      develop a new FISH protocol for rapid and specific detection of Leg. pneumophila 
      in water samples. METHODS AND RESULTS: A novel catalysed reporter deposition FISH 
      (CARD-FISH) protocol for the detection of Leg. pneumophila was developed, which 
      significantly enhanced signal intensity as well as specificity of the probe 
      through the use of a novel competitor probe. The developed protocol was compared 
      with the culture method for monitoring the seasonal development of culturable and 
      nonculturable Leg. pneumophila in two hospital cooling tower systems. Seasonal 
      fluctuations of Leg. pneumophila concentrations detected via CARD-FISH were 
      related to the development of the total bacterial community in both cooling 
      towers, with temperature and biocide as the main factors controlling this 
      development. CONCLUSIONS: Our results clearly showed that the majority of the 
      Leg. pneumophila cells were in a nonculturable state. Thus, detection of Leg. 
      pneumophila with culture methods may underestimate the total numbers of Leg. 
      pneumophila present. SIGNIFICANCE AND IMPACT OF THE STUDY: Rapid, sensitive and 
      specific detection and quantification of Leg. pneumophila in water systems is 
      prerequisite for reliable risk estimation. The new protocol significantly 
      improves current methodology and can be used to monitor and screen for Leg. 
      pneumophila concentrations in cooling towers or other water systems.
CI  - (c) 2012 The Authors. Journal of Applied Microbiology (c) 2012 The Society for 
      Applied Microbiology.
FAU - Kirschner, A K T
AU  - Kirschner AK
AD  - Institute for Hygiene and Applied Immunology, Water Hygiene, Medical University 
      Vienna, Vienna, Austria. alexander.kirschner@meduniwien.ac.at
FAU - Rameder, A
AU  - Rameder A
FAU - Schrammel, B
AU  - Schrammel B
FAU - Indra, A
AU  - Indra A
FAU - Farnleitner, A H
AU  - Farnleitner AH
FAU - Sommer, R
AU  - Sommer R
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120430
PL  - England
TA  - J Appl Microbiol
JT  - Journal of applied microbiology
JID - 9706280
RN  - 0 (Aerosols)
SB  - IM
MH  - Aerosols
MH  - Animals
MH  - *Environmental Microbiology
MH  - *Hospitals
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Legionella pneumophila/genetics/*isolation & purification
MH  - Legionnaires' Disease/microbiology/*prevention & control/transmission
MH  - Sensitivity and Specificity
MH  - Temperature
EDAT- 2012/03/27 06:00
MHDA- 2012/12/10 06:00
CRDT- 2012/03/27 06:00
PHST- 2012/03/27 06:00 [entrez]
PHST- 2012/03/27 06:00 [pubmed]
PHST- 2012/12/10 06:00 [medline]
AID - 10.1111/j.1365-2672.2012.05289.x [doi]
PST - ppublish
SO  - J Appl Microbiol. 2012 Jun;112(6):1244-56. doi: 10.1111/j.1365-2672.2012.05289.x. 
      Epub 2012 Apr 30.