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Related Experiment Videos

Real-time PCR method to detect Enterococcus faecalis in water.

Jorge W Santo Domingo1, Shawn C Siefring, Richard A Haugland

  • 1U.S. Environmental Protection Agency, National Exposure Research Laboratory, Microbial Exposure Research Branch, 26 W Martin Luther King Dr, Cincinnati, OH 45268, USA. santodomingo.jorge@epa.gov

Biotechnology Letters
|July 29, 2003
PubMed
Summary

A rapid real-time PCR method accurately detects Enterococcus faecalis in water. This technique identifies the bacteria in biofilms and freshwater samples in under three hours without nucleic acid extraction.

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Area of Science:

  • Environmental microbiology
  • Molecular biology
  • Water quality assessment

Background:

  • Enterococcus faecalis is an opportunistic pathogen found in water systems.
  • Accurate and rapid detection methods are crucial for public health.
  • Current detection methods can be time-consuming and complex.

Purpose of the Study:

  • To develop a sensitive and rapid 16S rDNA real-time PCR assay for Enterococcus faecalis detection.
  • To validate the assay's performance in various water matrices, including biofilms.
  • To establish a nucleic acid extraction-free method for faster results.

Main Methods:

  • Development of a 16S rDNA real-time PCR assay.
  • Quantification of Enterococcus faecalis using a dynamic range spanning five logs.

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  • Determination of the assay's limit of detection (LOD) at 6 CFU/reaction.
  • Testing the assay with spiked freshwater samples and biofilms from a water distribution system simulator.

    • Main Results:

    • The real-time PCR assay demonstrated a wide dynamic range for cell detection.
    • The limit of detection was as low as 6 CFU/reaction, indicating high sensitivity.
    • Enterococcus faecalis was successfully detected in both freshwater samples and complex biofilm matrices.
    • The assay provided results in less than 3 hours, eliminating the need for nucleic acid extraction.

    Conclusions:

    • The developed 16S rDNA real-time PCR assay is a sensitive, rapid, and effective tool for detecting Enterococcus faecalis in water.
    • The assay's ability to detect bacteria in biofilms and freshwater without prior nucleic acid extraction offers significant advantages for water quality monitoring.
    • This method holds promise for improved surveillance and management of waterborne pathogens.