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Related Experiment Videos

Quantitative real-time Legionella PCR for environmental water samples: data interpretation.

Philippe Joly1, Pierre-Alain Falconnet, Janine André

  • 1Centre National de Référence des Legionella, INSERM E-0230, Faculté de Médecine, IFR 62, 7 rue Guillaume-Paradin, 69372 Lyon Cedex 08, France.

Applied and Environmental Microbiology
|April 7, 2006
PubMed
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Quantitative PCR for Legionella in hot water systems shows promise, but laboratory-specific cutoffs are needed for accurate results. This method is less reliable for cooling tower water samples.

Area of Science:

  • Environmental microbiology
  • Molecular diagnostics

Background:

  • Legionella bacteria are a significant cause of Legionnaires' disease, often found in water systems.
  • Accurate detection of Legionella in water is crucial for public health.
  • Quantitative Polymerase Chain Reaction (qPCR) offers a rapid alternative to traditional culture methods.

Purpose of the Study:

  • To evaluate the performance of quantitative Legionella PCR assays in detecting Legionella in hot water systems and cooling towers.
  • To compare qPCR results with conventional culture methods.
  • To determine the applicability and limitations of qPCR for routine environmental monitoring.

Main Methods:

  • Quantitative PCR targeting the 16S rRNA gene (genus-specific) and mip gene (Legionella pneumophila-specific) were used.

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  • Samples included 223 hot water systems and 37 cooling tower water samples.
  • PCR results were compared against standard bacterial culture counts (CFU/liter).

    • Main Results:

    • 16S rRNA gene qPCR was nonquantifiable in up to 39.1% of hot water samples, predicting low Legionella counts (<250 CFU/liter).
    • Laboratory-specific qPCR cutoffs for hot water systems showed high sensitivity (87.7–92.9%) and specificity (77.3–96.5%), with mip PCR yielding the best specificity.
    • qPCR cutoffs were not reliably determined for cooling tower samples due to high variability and frequent culture-negative, PCR-positive results.

    Conclusions:

    • Quantitative Legionella PCR is applicable for monitoring hot water systems.
    • Positivity cutoffs for qPCR in hot water systems must be established independently by each laboratory.
    • qPCR is currently less reliable for Legionella detection in cooling tower water samples compared to hot water systems.