Background: Urinary tract infection (UTI) in neonates (infants ≤28 days of age) is associated with bacteremia and congenital anomalies of the kidney and urinary tract. Upper tract infections (acute pyelonephritis) may result in renal parenchymal scarring and chronic kidney disease. Neonates with UTI should be evaluated for associated systemic infection, and anatomic or functional abnormalities of the kidneys and urinary tract. The aim of our study is to detect the prevalence of urinary tract infection in listless neonates. Patients and methods: Across sectional study was carried out looked for urinary tract infection among sample of listless neonates in Bint Al Huda teaching hospital for maternity and children at Nasiriya city, Iraq at period from first of February to first of August, 2024. Information were gathered according to data collecting sheet that included: name, age, (gestational and postnatal), sex, mode of delivery and symptoms like lethargy, poor feeding, fever, shortness of breath and apneic attack. ect. Data have been obtained by standard laboratory procedures including GUE and urine culture. Result: A total of 100 neonates were enrolled in this study, the mean age of sample is (8.6 days), the minimum is 1 day and the maximum is 28 days, 75 were males and 25 were females. The percentage of urinary tract infection among listless neonates with a percentage of (75.8%), (24.2%) in males and females respectively. The current study found that age more than 7 days was significantly risk factor for urinary tract infection in addition to urinary tract infection during pregnancy among mothers both are significant risk factor (P value less than 0.001) from other point of view neonates of male gender, poor antenatal care was more likely to have urinary tract infection, however, the correlation with these possible risk factors did not reach the statistical significance (P value more than 0.001). It could be concluded from this study that UTI is a significant cause for lethargy. Urine culture may be used as an effective procedure to diagnose UTI, but to validate these finding larger studies are required.