Advertisement

Detection of Indicator Bacteria and Pathogens in Water by Polym Erase Chain Reaction (PCR) and Gene Probe Methods

  • R. M. Atlas
  • A. Bej
  • M. Mahbubani
  • R. Steffan
  • M. Perlin
  • J. DiCesare
  • L. Haff

Abstract

Sensitive methods for detecting coliform bacteria, and pathogenic bacteria (Salmonella and Shiqella) in environmental waters that do not require culturing of bacteria were developed by using the polymerase chain reaction (PCR) and gene probes. Cells were collected by filtration and DNA was released by freeze-thaw cycling. PCR amplification of region of lacZ gene was used as a target for detection of total coliform bacteria. A region of the lamB gene was the target for detection of Escherichia coli, Salmonella and Shigella. A region of the uid gene was used for detection of E. coli and Shigella. Biotin was incorporated during PCR amplification and the biotinylated DNA was detected by using membrane-bound poly-T-tailed capture probes. As little as 1 fg of total DNA and as few as 1 cell per 100 ml were detectable by the PCR-gene probe method.

Keywords

Gene Probe Fecal Coliform Capture Probe Total Coliform Fecal Contamination 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Greenberg, H., Ed., “Standard Methods for the Examination of Water and Wastewater,” American Public Health Association, Washington, D.C., 1985.Google Scholar
  2. 2.
    Edberg, S.C. and Edberg, M.M., “A defined Substrate Technology for the Enumeration of Microbial Indicators of Environmental Pollution,” Yale Journal of Biological Medicine, 61,389–399, 1988.Google Scholar
  3. 3.
    Edberg, S.C, Allen, M.J., Smith, D.B., and the national collaborative study, “National Field Evaluation of a Defined Substrate Method for the Simultaneous Detection of Total Conforms and Escherichia coli from Drinking Water: Comparison with Presence-absence Techniques,” Applied and Environmental Microbiology, 55,1003–1008,1989.PubMedGoogle Scholar
  4. 4.
    Mahbubani, M.H., Bej, A.K., Miller, R., Haff, L., DiCesare, J., and Atlas, R.M., “Detection of Legionella by Using Polymerase Chain Reaction and Gene Probe Methods”, Molecular and Cellular Probes, 4, 175–187, 1990.Google Scholar
  5. 5.
    Atlas, R.M., Bej, A.K., McCarty, S., DiCesare, J., and Haff, L., “Monitoring Microbial Pathogens and Indicator Microorganisms in Water by Using Polymerase Chain Reaction and Gene Probes”, Monitoring Water in the 1990’s: Meeting New Challenges, ASTM STP 1102, Jack R. Hall, G. Douglas Glysson, ed., American Society for Testing and Materials, Philadelphia, 1991.Google Scholar
  6. 6.
    Bej, A.K., Steffan, R.J., DiCesare, J., Haff, L., and Atlas, R.M., “Detection of Coliform Bacteria in Water by Polymerase Chain Reaction and Gene Probes”, Applied and Environmental Microbiology, 56, 306–314, 1990.Google Scholar
  7. 7.
    Bej, A.K., Mahbubani, M.H., Miller, R., DiCesare, J.L, Haff, L., and Atlas, R.M., “Multiplex PCR Amplification and Immobilized Capture Probes for Detection of Bacterial Pathogens and Indicators in Water”, Molecular and Cellular Probes, 4, 353–365, 1990.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1992

Authors and Affiliations

  • R. M. Atlas
  • A. Bej
    • 1
  • M. Mahbubani
    • 1
  • R. Steffan
    • 1
  • M. Perlin
    • 1
  • J. DiCesare
    • 1
    • 2
  • L. Haff
    • 1
    • 2
  1. 1.Dept. of BiologyUniv. of LouisvilleUSA
  2. 2.Perkin-Elmer Corp.NorwalkUSA

Personalised recommendations