Non-culturable Legionella pneumophila associated with Acanthamoeba castellanii: detection of the bacterium using DNA amplification and hybridization.
The intracellular localization of Legionella pneumophila serogroup 1 within Acanthamoeba castellanii rendered the bacteria non-culturable on supplemented BCYE agar. DNA amplification, using two 19-mer primers, and hybridization using a 25-mer oligonucleotide probe, permitted detection of Leg. pneumophila in approximately 81% (29/36) of samples where the bacteria could not be detected using culture. A combination of co-cultivation of samples with Leg. pneumophila-naive A. polyphaga or Hartmannella vermiformis, incubation in a defined liquid medium or use of catalase indicated that approximately 31% (9/29) of the samples contained Leg. pneumophila which were viable although not culturable.
Hay
J
J
Department of Ophthalmology, University of Glasgow, UK.
Seal
D V
DV
Billcliffe
B
B
Freer
J H
JH
D000048:Acanthamoeba / Q000382:microbiology*; D000818:Animals; D001431:Bacteriological Techniques; D003470:Culture Media; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D006249:Hartmannella / Q000382:microbiology; D016952:Legionella pneumophila / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction
D003470:Culture Media; D004269:DNA, Bacterial