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Specific detection of Escherichia coli isolated from water samples using polymerase chain reaction targeting four genes: cytochrome bd complex, lactose permease, β‐ d ‐glucuronidase, and β‐ d ‐galactosidase
Author(s) -
Horakova K.,
Mlejnkova H.,
Mlejnek P.
Publication year - 2008
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2008.03838.x
Subject(s) - escherichia coli , biology , permease , lactose permease , cytochrome , multiplex polymerase chain reaction , shigella , polymerase chain reaction , gene , microbiology and biotechnology , enterobacteriaceae , bacteria , biochemistry , enzyme , genetics
Aims: To develop a PCR‐based method for reliable detection of Escherichia coli that enables its differentiation from biochemically and phylogenetically related bacteria. Methods and Results: Using multiplex PCR targeting four genes (cytochrome bd complex, lactose permease, β‐ d ‐glucuronidase, and β‐ d ‐galactosidase) the possibility of specific detection of various control E. coli strains was tested. It was found that four PCR fragments of the predicted size were observed only for E. coli strains, but not for relatives as close as Shigella sp. or other enterobacteria. Not surprisingly, this method enabled us to identify also E. coli strains which did not exhibit the β‐ d ‐glucuronidase activity. Our multiplex PCR was also successfully used for identification of 95 environmental isolates of E. coli . Conclusions: The developed PCR‐based method, in which four genes coding for lactose permease, cytochrome bd complex, β‐ d ‐glucuronidase, and β‐ d ‐galactosidase, serve as target DNA sequences, allows precise and reliable detection of E. coli strains. Significance and Impact of the study: The suggested approach increases the specificity of detection of E. coli since it enables to distinguish E. coli from Shigella sp. and other relative enterobacteria.