Skip to main content
SpringerLink
Log in

Real-time PCR method to detect Enterococcus faecalis in water

  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

A 16S rDNA real-time PCR method was developed to detect Enterococcus faecalis in water samples. The dynamic range for cell detection spanned five logs and the detection limit was determined to be 6 cfu/reaction. The assay was capable of detecting E. faecalis cells added to biofilms from a simulator of a water distribution system and in freshwater samples. Nucleic acid extraction was not required, permitting the detection of E. faecalis cells in less than 3 h.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Belgrader P, Benett W, Hadley D, Richards J, Stratton P, Mariella R, Milanovich F (1999) PCR detection of bacteria in seven minutes. Science 284: 449–450.

    Google Scholar 

  • Cox T, Frazier C, Tuttle J, Flood S, Yagi L, Yamashiro CT, Behari R, Paszko C, Cano RJ (1998) Rapid detection of Listeria monocytogenes in dairy samples utilizing a PCR-based fluorogenic 5´ nuclease assay. J. Ind. Microbiol. Biotechnol. 21: 167–174.

    Google Scholar 

  • Devriese LA, Laurier L, De-Herdt P, Haesebrouck F (1992) Enterococcal and streptococcal species isolated from feaces of calves young cattle and dairy cows. J. Appl. Bacteriol. 72: 29–31.

    Google Scholar 

  • Dufour A, Ballentine R (1986) Ambient water quality criteria for bacteria. EPA 440-5-84-002. Office of Research and Development, U.S. Environmental Protection Agency, Washington, D.C.

    Google Scholar 

  • Haugland R, Vesper SJ, Wymer LJ (1999) Quantitative measurement of Stachybotrys chartarum conidia using real time detection of PCR products with the TaqMan (TM) fluorogenic probe system. Mol. Cell. Probes 13: 329–340.

    Google Scholar 

  • Maidak BL, Cole JR, Lilburn TG, Parker CT, Saxman PR, Stredwick JM, Garrity GM, Li B, Olsen GJ, Pramanik S, Schmidt TM, Tiedje JM (2000) The RDP (Ribosomal Database Project) continues. Nucl. Acids Res. 28: 173–174.

    Google Scholar 

  • Messer JW, Dufour AP (1998) A rapid, specific membrane filtration procedure for enumeration of enterococci in recreational water. Appl. Environ. Microbiol. 64: 678–680.

    Google Scholar 

  • Nelson RR, McGregor KF, Brown AR, Amyes SG, Young H (2000) Isolation and characterization of glycopeptide-resistant enterococci from hospitalized patients over a 30-month period. J. Clin. Microbiol. 38: 2112–2116.

    Google Scholar 

  • Oberst RD, Hays MP, Bohra LK, Phebus RK, Yamashiro CT, Paszko-Kolva C, Flood SJA, Sargeant JM, Gillespie JR (1998) PCR-based DNA amplification and presumptive detection of Escherichia coli O157:H7 with an internal fluorogenic probe and the 5´ nuclease (TaqMan) assay. Appl. Environ. Microbiol. 64: 3389–3396.

    Google Scholar 

  • Patel R, Piper KE, Rouse MS, Steckelberg JM, Uhl JR, Kohner P, Hopkins MK, Cockerill FR, Kline BC (1998) Determination of 16S rRNA sequences of enterococci and application to species identification of nonmotile Enterococcus gallinarum isolates. J Clin. Microbiol. 36: 3399–3407.

    Google Scholar 

  • Pourcher AM, Devriese LA, Hernandez JF, Delattre (1991) Enumeration by a miniaturized method of Escherichia coli, Streptococcus bovis, and enterococci as indicators of the origin of fecal pollution of waters. J. Appl. Bacteriol. 70: 525–530.

    Google Scholar 

  • Reasoner DJ, Geldreich EE (1985) A new medium for the enumeration and subculture of bacteria from potable water. Appl. Environ. Microbiol. 49: 1–7.

    Google Scholar 

  • Sen K (2000) Rapid identification of Yersinia enterocolitica in blood by the 5´ nuclease PCR assay. J. Clin. Microbiol. 38: 1953–1958.

    Google Scholar 

  • Wang RF, Cao WW, Cerniglia CE (1996) PCR detection and quantitation of predominant anaerobic bacteria in human and animal fecal samples. Appl. Environ. Microbiol. 62: 1242–1247.

    Google Scholar 

  • Wheeler AL, Hartel PG, Godfrey DG, Hill JL, Segards WI (2002) Potential of Enterococcus faecalis as a human fecal indicator for microbial source tracking. J. Environ. Qual. 31: 1286–1293.

    Google Scholar 

Download references

Author information

Author notes

  1. Jorge W. Santo Domingo

    Present address: U.S. Environmental Protection Agency, National Risk Management Research Laboratory, Water, MS-387, 26 W. Martin Luther King Dr., Cincinnati, OH, 45268, USA

Authors and Affiliations

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

    Jorge W. Santo Domingo, Shawn C. Siefring & Richard A. Haugland

Authors
  1. Jorge W. Santo Domingo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shawn C. Siefring
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Richard A. Haugland
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jorge W. Santo Domingo.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Santo Domingo, J.W., Siefring, S.C. & Haugland, R.A. Real-time PCR method to detect Enterococcus faecalis in water. Biotechnology Letters 25, 261–265 (2003). https://doi.org/10.1023/A:1022303118122

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1022303118122

Search

Navigation