Studies on the Detection of Escherichia coli and Coliform Bacteria in Water, Meat and Vegetables by the Polymerase Chain Reaction
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We applied the polymerase chain reaction (PCR) to the detection of <I>Escherichia coli</I> and coliform bacteria in water and food. Primer sets targeting the transferase gene (<I>wec</I>F), the β-galactosidase gene (<I>lacZ</I>) and the 16S rRNA gene (<I>E. coli</I> 16S rRNA) of <I>E. coli</I> were used in the PCR, and the following results were obtained.<BR>1) The frequencies of preservation of the <I>wec</I>F, <I>lac</I>Z and <I>E. coli</I> 16S rRNA genes were 12/12 (100%), 99/99 (100%) and 99/99 (100%), respectively, for <I>E. coli</I> from food and human feces, and 28/63 (44%), 70/177 (39%) and 0/177 (0%), respectively, for coliform bacteria including <I>Klebsiella</I> spp., <I>Citrobacter</I> spp. and others from food.<BR>2) The detection frequencies of the <I>wec</I>F, <I>lac</I> and <I>E. coli</I> 16S rRNA genes from the BGLB culture of 69 samples of food (23 samples of meat and 46 samples of vegetables) were 73%, 65% and 37%, respectively. A significant correlation was found between the isolation of <I>E. coli</I> and the detection of the <I>E. coli</I> 16S rRNA gene, but no correlation was found between the isolation of coliform bacteria and the detection of the <I>E . coli</I> 16S rRNA gene. Also, no significant correlation was found between the isolation of coliform bacteria and the detection of the <I>wec</I>F or <I>lacZ</I>gene.<BR>3) A significant correlation between the detection of <I>E. coli</I> 16S rRNA and the isolation of <I>E. coli</I> was observed, when fecal contamination of 153 water samples from various sources of water supply were examined by the defined substrate method using colilert.<BR>These results show that PCR targeting the <I>E. coli</I> 16S rRNA gene is useful for rapid and specific detection of <I>E. coli</I> in water and food.
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