STP1102: Monitoring Microbial Pathogens and Indicator Microorganisms in Water by Using the Polymerase Chain Reaction and Gene Probes

    Atlas, RM
    professor of Biologyresearch scientistresearch scientistresearch scientists, the University of Louisvillethe University of Louisvillethe Louisville Water CompanyPerkin-Elmer Corp., LouisvilleLouisvilleLouisvilleNorwalk, KYKYKYCT

    Bej, AK
    professor of Biologyresearch scientistresearch scientistresearch scientists, the University of Louisvillethe University of Louisvillethe Louisville Water CompanyPerkin-Elmer Corp., LouisvilleLouisvilleLouisvilleNorwalk, KYKYKYCT

    McCarty, S
    professor of Biologyresearch scientistresearch scientistresearch scientists, the University of Louisvillethe University of Louisvillethe Louisville Water CompanyPerkin-Elmer Corp., LouisvilleLouisvilleLouisvilleNorwalk, KYKYKYCT

    DiCesare, J
    professor of Biologyresearch scientistresearch scientistresearch scientists, the University of Louisvillethe University of Louisvillethe Louisville Water CompanyPerkin-Elmer Corp., LouisvilleLouisvilleLouisvilleNorwalk, KYKYKYCT

    Haff, L
    professor of Biologyresearch scientistresearch scientistresearch scientists, the University of Louisvillethe University of Louisvillethe Louisville Water CompanyPerkin-Elmer Corp., LouisvilleLouisvilleLouisvilleNorwalk, KYKYKYCT

    Pages: 10    Published: Jan 1991


    Abstract

    A method using the polymerase chain reaction (PCR) and gene probes was developed for the detection of enteric pathogens and indicators of fecal contamination in waters. The method involves collection of cells from 100 mL water samples by filtration, lysis of cells, PCR amplification of regions of the lacZ and lamB genes and detection of the amplified target DNA by gene probes. Using lacZ as the target gene “total coliform bacteria” were detected. Using lamB as the target gene the fecal indicator Escherichia coliand the enteric pathogens Shigella and Salmonella were specifically detected. The sensitivity of detection was 1–5 viable target microorganisms per 100 mL sample and the test procedure takes only a few hours. Results with PCR-gene probe tests were found to be comparable to those of conventional plating and the Colilert procedures. The PCR-gene probe method for coliform detection can be used as an effective monitoring procedure for insuring the bacteriological quality of water supplies.

    Keywords:

    PCR, gene probe, coliforms, water quality, indicator bacteria


    Paper ID: STP17615S

    Committee/Subcommittee: D19.24

    DOI: 10.1520/STP17615S


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