Introduction and Objectives: Urinary tract infections are among the most prevalent inflectional diseases. The phylogenetic analysis showed that the strains of Escherichia coli (E.coli) are classified into four different groups, A1, B1, B2, and D. The aim of this research was to determine the phylogenetic groups of E.coli isolated from urine infections in Mashhad City. Materials and Methods: This study was conducted on 111 E. coli isolates from the urine culture of the subjects inflicted with urinary tract infections (69 isolates from outpatients and 42 isolates from the inpatients). The biochemistry and microbial tests were employed to verify the isolates’ inspections. A method has been used to identify the phylogenetic groups, based on Multiplex PCR, using a combination of two genes, including Chua gene and YjaA gene and a single piece of DNA, TapE4.C2. Results: The most prevalent phylogenetic group among 69 E. coli isolates from the outpatients belonged to D2 (%32) and (%24.6). Also, the most prevalent phylogenetic group among all the E. coli isolates from inpatients belonged to B23 (%57.1), A1 (%16.7), D2 (%14.3), and B22 (%7.1). Among all E. coli isolates from inpatients and outpatients, 2 and 4 isolates could not be classified, through Clermont method, respectively. Conclusion: The results of statistical analysis showed that B23 phylogenetic group has been significantly higher for inpatients, compared to the outpatients and D2 phylogenetic group versus. The difference in the distribution of other phylogenetic groups of E. coli has not been significant for inpatients and outpatients. Further, among the inpatients, no significant difference regarding age, gender, and phylogenetic group was observed.

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