A total of 100 samples were collected from patients suffering from urinary tract infections were admitted and visited Maternity and Children Hospital in Al-Hilla city, during a period of three months from September to November 2021. To confirm the identification of E. coli by use selective media, biochemical tests, Api20E system and automated Vitek 2 system, it was found that, 33 isolates were showed positive and identified as E. coli. Molecular detection of some virulence factors genes were studied, out of 33 E. coli isolates, fimH gene was detected in 27(81.8%) isolates by the presence of (508 bp), pap gene was detected in 26(78.8%) isolates by the presence of (336bp), hlyA gene was detected in 10(30.3%) isolates by the presence of (1177bp), feoB gene was detected in 19(57.6%) isolates by the presence of (470bp), usp gene was detected in 21(63.6%%) isolates by the presence of (440bp) and 24(72.7%) have iroN gene by the presence of (665bp) compared with allelic ladder.