Development of PCR protocol for detection of Escherichia ... - WIT Press
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Development of PCR protocol for detection of Escherichia ... - WIT Press
Reaction (PCR) is a molecular method, which can be used for detection of ... The primer pairs used for Escherichia coli O157:H7 detection were Rfb-F, Rfb-.
Water Resources Management VII
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Development of PCR protocol for detection of Escherichia coli in drinking water J. Imtiaz, I. Hashmi, A. Saeed, I. A. Qazi & M. Arshad Institute of Environmental Sciences and Engineering (IESE), School of Civil and Environmental Engineering (SCEE), National University of Science and Technology (NUST), Pakistan
Abstract 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 (
