POLYMERASE CHAIN REACTION ASSAY FOR DETECTION OF ENTEROPATHOGENIC ESCHERICHIA COLI STRAINS IN MEAT
2011
Stanilova, S., Thracian University, Stara Zagora (Bulgaria) | Rusenova, N., Thracian University, Stara Zagora (Bulgaria) | Petrova, D., Thracian University, Stara Zagora (Bulgaria) | Stoyanchev, T., Thracian University, Stara Zagora (Bulgaria)
Enteropathogenic Escherichia coli - EPEC strains are a significant cause of acute and persistent diarrhea in humans, which are transmitted primarily through consumption of contaminated foods. The present study was performed for the identification of EPEC strains from contaminated meat by a PCR technique. A panel of reference E. coli strains purchased from NBIMCC; Sofia, Bulgaria was used for the experiments. Using the optimized amplification conditions, PCR with the purified DNA template was amplified for eae gene and obtained amplicons were visualized by 2% agarose gel electrophoresis. Our result demonstrated that two strains: E. coli O 111-НБПМКК-4447 and E. coli O 26-НБПМКК-7434 have amplified a 482 bp fragment with used primers. In our experimental condition the lowest cell concentration which exhibit well visualized 482 bp amplification band is less then 54 viable E. coli cells in 1 ml. The lowest bacterial cell concentration - CFU/g in the contaminated beef meat samples which was detected in our experiments was 0,7 CFU/g meat. In conclusion the developed PCR method could be used for detection of typical and atypical EPEC in isolated colonies and in food.
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