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Updated: Aug 10, 2026

Purification of Pathogen Vacuoles from Legionella-infected Phagocytes
Published on: June 19, 2012
Non-culturable Legionella pneumophila associated with Acanthamoeba castellanii: detection of the bacterium using DNA
J Hay1, D V Seal, B Billcliffe
1Department of Ophthalmology, University of Glasgow, UK.
Abstract:
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.

