Background: Urinary tract infection is the most common site of infection associated with health care in the Intensive Care Unit (ICU) patients. Long-term catheterization is the most important risk factor for Urinary Tract Infections (UTIs). These infections are more common in women than in men. Colonized microorganisms in the perineal skin and around the urethra may move into the urinary system through the external surfaces of the urinary catheter to cause infection. Objectives: The present study aimed to compare the effect of perineal care with normal saline and 2% chlorhexidine solution on the rate of catheter-associated urinary tract infection (CAUTI) in women hospitalized in ICUs. Methods: A quasi-experimental study was performed on 70 female patients hospitalized in the ICU of Khatam-Al-Anbia Hospital of Zahedan in 2019. The research units meeting the inclusion criteria were selected using convenience sampling and then randomly divided into two groups of normal saline (n = 35) and chlorhexidine (n = 35). The perineal area of patients in the first group was washed with normal saline and the second group with 2% chlorhexidine solutions twice a day for seven days in both groups. To diagnose UTIs, urine samples were cultured on the first and seventh days. Data were analyzed using SPSS 21 via the independent t-test, chi-square test, and Fisher’s exact test at a significance level of less than 0.05. Results: Finally, 60 participants completed the study. The two groups did not differ significantly in terms of age, marital status, level of education, cause of hospitalization, antibiotic use, underlying diseases, fluid intake, and urinary output. At the end of the seventh day after the intervention, the incidence of UTIs was significantly lower in the chlorhexidine group (13.3%) than in the normal saline group (76.7%) (P = 0.001). Conclusions: The results showed that washing the perineal area every 12 hours with 2% chlorhexidine solution compared to normal saline reduces the incidence of CAUTIs in women hospitalized in ICUs. Therefore, perineal care with a 2% chlorhexidine solution is recommended for perineal care of female patients with urinary catheters hospitalized in ICUs.