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This work was to genetically characterized Shiga toxin-producing Escherichia coli strains isolated from imported beef meat in Malaysia using polymerase chain reaction (PCR) base analysis, seventy four (74) frozen beef meats samples (imported) (n=74) tested originated forty two of the sample were from India and thirty two of the samples were from Australia. The samples were bought from the frozen meat units of five different supermarkets in diverse settings in Malaysia which start from 2012 April to 2013 October on a weekly basis. E. coli and shiga toxin producing E. coli isolation was done Isolation of plasmid was carried out using the PureYield™ Plasmid Miniprep System. Enterobacterial Repetitive Intergenic Consensus (ERIC) primer was use in genetically isolating the bacteria: We used seven types of primers for Random Amplified Polymorphic DNA-PCR (RAPD-PCR) namely Gen8, Gen9, A1, A7, A10, OPAR8 and OPAR20. Our result revealed that Plasmid profiling showed 16 patterns based on size and 4 patterns on gel. Combination of phenotypically and genotypically approaches revealed a varied heterogeneity among imported beef isolates of E. coli. Isolated plasmids from shiga toxin producing E. coli varied in sizes. The sizes are from 4.3 -23.1 kilo base (kb). Sixty two (62) isolates were found to harbor plasmid with size 23.2 kb while 43 isolates harbored more than a single plasmid. The analysis of data through the use of average linkage (UPGMA, unweight group pair technique with arithmetic averages) using Gel compare 11 software which was displayed in dendrograms, for the, ERIC-PCR and RAPD-PCR (RAPD Gen 8, Gen 9), (Opar 8 and Opar 20), (RAPD A1, RAPD A7and RAPD A10) analysis, produced various cluster. The PCR analysis using OPAR 8 and OPAR 20 also produces various clusters. |
