Molecular detection of Pseudomonas aeruginosa in recreational water

Int J Environ Health Res. 2012;22(1):60-70. doi: 10.1080/09603123.2011.588325. Epub 2011 Jun 13.

Abstract

The aim of this study was the development of a new molecular assay for Pseudomonas aeruginosa identification in recreational water. The method includes bacterial cell concentration through membrane filtration, a short (6 h) culture-enrichment step, DNA extraction and its amplification through a Real-Time PCR assay. The performance of the molecular approach was evaluated on 44 samples of swimming pool water and compared with the reference method UNI EN ISO 16266:2008. Positivity rates of 6% and 74% in pool and inlet water, respectively, with the standard culture method, and of 23% and 74% with the molecular method were found. Statistical analysis indicated "substantial agreement" (Cohen's Kappa index: 0.6831) between the two approaches. RAPD typing of P. aeruginosa isolates showed identical fingerprint profiles, indicating their epidemiological correlation. The developed protocol showed very high specificity and a detection limit of 10 genomic units. This technique has the potential to screen large numbers of environmental samples, and could be proposed as part of a self-monitoring plan for recreational facilities, improving surveillance and early warning systems.

Publication types

  • Validation Study

MeSH terms

  • DNA, Bacterial / analysis*
  • DNA, Bacterial / genetics
  • Environmental Monitoring / methods*
  • Filtration
  • Membranes
  • Pseudomonas aeruginosa / genetics
  • Pseudomonas aeruginosa / isolation & purification*
  • Random Amplified Polymorphic DNA Technique
  • Real-Time Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Swimming Pools
  • Water Microbiology*

Substances

  • DNA, Bacterial