Objective: To study the distribution and drug resistance of pathogens in urinary tract infections(UTI), and to provide evidence for rational treatment of the disease. Methods: The results of bacterial culture from morning clean-catch mid-streamurinein the patients with UIT during July 2016 to June 2020 were retrospectively analyzed to determine the distribution and drug resistance of pathogens. Result A total of 3 543 specimens were collected, 810 strains of pathogens were isolated, and the positive rate was 22.9%, including 558 strains of Gram-negative bacteria (68.9%), 113 strains of Gram-positive cocci (14.0%), 128 strains of Mycoplasma(15.8%), 11 strains of fungi(1.4%). The majority of bacteria strains were Escherichia coli (337 strains, 41.6%) and Klebsiella pneumoniae (67 strains, 8.3%). Ureaplasma urealyticum(UU) accounted for 10.6% (86 strains) inisolated mycoplasma. Escherichia coli and Proteusisolates were sensitive to aminoglycoside and carbapenem antibiotics and the resistant rates were lower than 5%; Klebsiella pneumoniae was resistant to ceftriaxone and levofloxacin and its′ resistant rates were 58.3% and 55.2%, respectively.All Enterococci strains were resistant to penicillin, ampicillin, gentamicin, and levofloxacin, which had higher resistance rates than those in Enterococcus faecalis; Enterococcus faecalis had high resistant rates to levofloxacin and moxifloxacin(73.3% and 64.4%, respectively). Staphylococcus aureus and Enterococcus were sensitive to sulfonamides, glycopeptides and common broad-spectrum antibiotics, and the resistant rates were lower than 5%. The drug resistant rate of UU to ciprofloxacin was 69.9%, and UU was sensitive to both minocycline and doxycycline. Conclusions: The majority of pathogens isolated from UTI patients in Zhoupu District of Shanghai is Gram-negative bacilli, which shows different degree of resistance to common antibiotics. Therefore, the regular monitoring of UTI pathogenic bacteria is important to determine how to choose antibiotics in clinical practice.