Aims: This study was conducted to isolate and identify by PCR analysis for presence stx1gene, stx2 gene and fliCh7 gene of Shiga toxin Escherichia coli from broiler chickens and humans from live chicken markets in different areas of Karbala. Material and methods: A total of 200 fecal samples were collected from chicken broiler (n=150) and human (n=50) . E. coli were isolated and identified by cultural, biochemical, Api-20E, VITEK 2 system and PCR. Results: 13 Human and chicken positive samples include 10% , 5.3% and were derived from human and broiler chickens respectively that diagnosis by using PCR analysis for presence stx1gene, stx2 gene and fliCh7 gene. Shiga toxin–producing E. coli results negative stx1gene and positive stx2 gene . Conclusion: Considering the findings of this study demonstrate that the chicken's digestive track hosts a bacterial pathogen, indicating that interferences are required to reduce Shiga-toxin-producing Escherichia coli(STEC) transmission.