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.
Tsen
H Y
HY
Department of Food Science, National Chung-Hsing University, Taichung, Taiwan, Republic of China.
Lin
C K
CK
Chi
W R
WR
D017931:DNA Primers; D004926:Escherichia coli / Q000235:genetics / Q000302:isolation & purification*; D005798:Genes, Bacterial; D016133:Polymerase Chain Reaction / Q000379:methods; D012329:RNA, Bacterial / Q000032:analysis*; D012336:RNA, Ribosomal, 16S; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014871:Water Microbiology
D017931:DNA Primers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S