Despite the fact that the pharmaceutical industry has developed numerous novel antibiotics over the last three decades, bacteria have developed resistance to these medications. Bacteria have the genetic potential to transfer and develop resistance to medications used as therapeutic agents. Therefore, the present study aimed to reveal the antibacterial role of Achillea millefolium against Escherichia coli isolated from urinary tract infections. Between February and July of 2024, 130 clinical samples were taken from individuals who were admitted to Kirkuk Hospital in Kirkuk City after visiting a specialist and being sent to the laboratory. Urine samples were taken from patients with UTI ranging in age from (1-60 years) of women, after which they were transferred directly to the laboratory to be cultured on the culture media, then A. millefolium extract was tested against Escherichia coli, compared with some standard antibiotics. Results showed that 57 (43.8%) of the total samples showed positive results for bacterial growth when cultured on the best culture media, which included blood and MacConkey agars. 73 (or 56.2%) of the total samples had results that showed no bacterial growth. E. coli showed total resistance (100%) to Ampicillin. While, a low sensitive toward Nalidixic acid (20.0%), Clindamycin (20.0%) 21.5%. Otherwise, E. coli showed a high sensitive to Gentamycin (87.7%), Imipenem (96.9%), and Amikacin (90.8%). While Tobromycin showed high sensitive 100.0% respectively. Based on the results, it was shown that the extract of A. millefolium at a dosage of 100 mg induced an inhibition diameter rate of 18.7±3.45, while at a concentration of 150 mg, it produced an inhibition diameter rate of 29.49±2.97. It is concluded from that the alcoholic extract of A. millefolium has an antibacterial effect and is very effective in treating cases of urinary tract infection resulting from infection with Escherichia coli.