Abstract
Polymerase chain reaction (PCR) amplification and gene probe detection of regions of two genes, lacZ and lamB, were tested for their abilities to detect coliform bacteria. Amplification of a segment of the coding region of Escherichia coli lacZ by using a PCR primer annealing temperature of 50 degrees C detected E. coli and other coliform bacteria (including Shigella spp.) but not Salmonella spp. and noncoliform bacteria. Amplification of a region of E. coli lamB by using a primer annealing temperature of 50 degrees C selectively detected E. coli and Salmonella and Shigella spp. PCR amplification and radiolabeled gene probes detected as little as 1 to 10 fg of genomic E. coli DNA and as a few as 1 to 5 viable E. coli cells in 100 ml of water. PCR amplification of lacZ and lamB provides a basis for a method to detect indicators of fecal contamination of water, and amplification of lamB in particular permits detection of E. coli and enteric pathogens (Salmonella and Shigella spp.) with the necessary specificity and sensitivity for monitoring the bacteriological quality of water so as to ensure the safety of water supplies.
MeSH Terms
Base Sequence
Blotting, Southern
DNA Probes
DNA, Bacterial/analysis,genetics
Electrophoresis, Agar Gel
Enterobacteriaceae/genetics,isolation & purification
Gene Amplification
Humans
Molecular Sequence Data
Nucleic Acid Hybridization
Polymerase Chain Reaction
Predictive Value of Tests
Species Specificity
Water Microbiology
Chemicals
DNA Probes
DNA, Bacterial
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Bej A K
Department of Biology, University of Louisville, Kentucky 40292.
Steffan R J
DiCesare J
Haff L
Atlas R M
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