Detection of coliform bacteria in water by polymerase chain reaction and gene probes.
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.
Bej
A K
AK
Department of Biology, University of Louisville, Kentucky 40292.
Steffan
R J
RJ
DiCesare
J
J
Haff
L
L
Atlas
R M
RM
D001483:Base Sequence; D015139:Blotting, Southern; D015342:DNA Probes; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D004755:Enterobacteriaceae / Q000235:genetics / Q000302:isolation & purification*; D005784:Gene Amplification; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D013045:Species Specificity; D014871:Water Microbiology
D015342:DNA Probes; D004269:DNA, Bacterial