Application of pulsed-field gel electrophoresis to evaluate genetic diversity of Escherichia coli from the environment of a cattle-rearing area and clean river system

Ng, Ling Nee (2006) Application of pulsed-field gel electrophoresis to evaluate genetic diversity of Escherichia coli from the environment of a cattle-rearing area and clean river system. [Final Year Project Report] (Unpublished)

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Abstract

Pulse-field gel electrophoresis (PFGE) had been applied widely to study the epidemiology of pathogenic Escherichia coli but its capability as a tool to explore the genetic relationship among E. coli isolates in Malaysia had not yet been evaluated fully. In this study, the genomic DNA from 32 Escherichia coli isolates obtained from cattle feces at Cattle rearing areas and its Secondary environments were subjected to PFGE after digestion with restriction enzyme Xbal. This was done in order to access the relationship of E coli isolated from the host to secondary environments and their possible transmission. PFGE results showed that E. coli isolates were successfully categorized to their sources and there was no clear correlation between isolates of host (primary) with secondary environment. Hence it was suggested that E. coli in secondary environment may have originated from other sources. In addition, a multiplex PCR using primers SLTI genes, SLTII genes, Rfb genes and FLICh -7 genes was done to detect the presence of pathogenic E.coli strains. All isolates (n = 31) did not possess the SLTI and SLTII genes, thus that PFGE was a useful method to evaluate the genetic diversity of £. coli from different sources. Besides, multiplex PCR was also proven to be a reliable tool for the identification of pathogenic E.coli strains.

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