Streptococcus pneumoniae is a leading cause of respiratory tract infections, including pneumonia. In recent years, due to the incidence of resistance to commonly used antibiotics, quinolones have been taken into consideration in treatment of S. pneumoniae. The present study aimed to investigate the quinolone resistance rate among pneumococcus isolates from Shiraz. This cross-sectional-descriptive study was performed on 45 strains of S. pneumoniae isolated from clinical samples. Initially, using biochemical tests and PCR technique with specific primers of lytA gene, S. pneumoniae strains were identified. Subsequently, antibiotic resistance to quinolones was evaluated by CLSI criteria. Among 45 strains isolated from clinical specimen, resistance to nalidixic acid, ciprofloxacin, ofloxacin, norfloxacin and levofloxacin were estimated 88.22, 73.43, 53.33, 48.88 and 42.22 %, respectively. The highest frequency of resistance corresponded to the people with pneumonia. Besides, the greatest resistance to all antibiotics related to the age group of 31–40. Statistical analysis indicated no significant association between antibiotic resistance and disease type or age groups. In concurrence with other investigations, our results showed that the level of resistance to quinolones is increasing in Shiraz. Hence, the quinolone resistance in this region is a serious issue; therefore, the regional pattern of antimicrobial resistance on therapeutic regimes must be taken into consideration.