Development and use of 16S rRNA gene targeted PCR primers for the identification of Escherichia coli cells in water

The primary sequences of the V₃ and V₆ regions of the 16S rRNA gene of pathogenic and non-pathogenic strains of Escherichia coli were determined and compared with those obtained for a number of reference strains which belong to the family Enterobacteriaceae. Three oligonucleotide primers 16E1, 16E2 and 16E3 were designed and used in the polymerase chain reaction to identify specifically all E. coli isolates. When 16E1, 16E2 and 16E3 were used as primers for the identification of E. coli cells present in tap, underground and pond waters, as low as 1 cfu 100 ml⁻¹ of water could be detected if an 8 h pre-culture step was performed prior to the PCR reaction.