Development Of PCR Protocol For Detection Of Escherichia Coli In Drinking Water

Escherichia coli is a pathogenic microorganism that may cause severe gastrointestinal illness in humans. This pathogen may be transmitted in a variety of ways, including food and water. As with most waterborne pathogens E. coli is difficult to detect and enumerate with accuracy in drinking waters du...

Full description

Saved in:
Bibliographic Details
Published in:WIT Transactions on Ecology and the Environment 2013-01, Vol.171, p.225
Main Authors: Imtiaz, J, Hashmi, I, Saeed, A, Qazi, I A, Arshad, M
Format: Article
Language:English
Subjects:
Bacteria
Coliforms
Contamination
Culture techniques
Deoxyribonucleic acid
DNA
Drinking water
E coli
Electrophoresis
Enrichment
Environmental monitoring
Escherichia coli
Ethidium bromide
Food contamination
Gel electrophoresis
Oligonucleotides
Optimization
Pathogens
Polymerase chain reaction
Primers
Purification
Toxins
Virulence
Water analysis
Water pollution
Water sampling
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Escherichia coli is a pathogenic microorganism that may cause severe gastrointestinal illness in humans. This pathogen may be transmitted in a variety of ways, including food and water. As with most waterborne pathogens E. coli is difficult to detect and enumerate with accuracy in drinking waters due to methodological limitations. The aim of this study was to develop a PCR protocol for the detection of E. coli O157:H7 and E. coli virulence gene SLT-I (Shiga like toxin) in drinking water using a double enrichment step. This method is comprised of bacterial DNA purification using Genomic DNA extraction kit followed by PCR detection. The PCR optimization was done with E. coli O157:H7 strain EDL 933 (ATCC 43895). The oligonucleotide primers Rfb and SLT-I were used for targeting O157 and SLT-I genes respectively. The specific PCR product of these primers were obtained at 292 bp and 210 bp for Rfb and SLT-I respectively, which were visualized by gel electrophoresis and ethidium bromide staining. In spiked water samples, PCR results showed high sensitivity (
ISSN:1746-448X
1743-3541
DOI:10.2495/WRM130201