Urinary tract infection (UTI) is a common infection in clinical practice, and the urine culture is regarded as the gold standard for final diagnosis and widely conducted in clinical microbiology laboratory. However, there are still some problems. These problems include whether it is necessary that all the urine specimens should be screened before culture, how to screen, under what conditions the urine specimens should be stained, how to understand correctly the clinical significance of the quantitative urine culture, how to understand the culture value of Corynebacterium urealyticum detected in UTI, and how to interpret the urinary culture results in acute cystitis in premenopausal women. The present review will discuss the above-mentioned issues in line with the studies in UTI, especially the recently-released health standards-the Diagnosis of Urinary Tract Infections in Clinical Microbiology Laboratory, recommended by National Health and Family Planning Commission of the People?s Republic of China.