Abstract: Objective: To evaluate the clinical effect of L-form bacterium culture in urine samples from patients with urinary infection. Methods: Five hundred and ten urine specimens were examined with the routine and L-form bacterium culture,the isolated strains were tested with drug susceptibility test. Results: Two hundred and thirty out of 510 urine specimens were identified as bacterial growth. Of them,100 wild-type strains and 60 L-form strains alone were isolated,and both the wild-type and the L-form strains from 70 urine samples. The drug susceptibility test were displayed that all of the L-form strains were susceptible to macrolides and tetracycline antibiotics,while the wild-type to penicllium and cephem antibiotics. Conclusions: Most of the urinary infection may be attributed to Lform bacterium. Therefore,the L-form bacterium culture should be done as a routine test and it contributes to the clinical diagnosis and rational use of antibiotics.