Urinary tract infection (UTI) is a common and costly disease that affects millions of people worldwide each year. Accurate laboratory diagnosis of UTI is crucial to reduce antimicrobial resistance in UTI pathogens because of unnecessary antimicrobial use. Routine urinalysis with urine dipstick analysis and Gram staining can be used to screen for UTI. The conventional ‘gold standard’ for diagnosing UTIs involves culture-based tests. This method entails culturing the urine sample to amplify the bacteria to detectable levels, followed by biochemical and serological tests, as well as antimicrobial susceptibility tests. However, culture-based tests have the disadvantage of requiring 48-72 hours to report results owing to the time needed for bacterial growth. Therefore, various methods have been developed and are used to diagnose UTI to replace time-consuming culture tests. These methods include flow cytometry, mass spectrometry, and nucleic-acid-based diagnostic tests. This review introduces various laboratory methods used to diagnose UTI in clinical microbiology laboratories and discusses their principles and interpretation methods.