Objective To investigate characteristics of clinical distribution and change in antimicrobial resistance of Group B Streptococcus(GBS), and provide scientific basis for the prevention and treatment of GBS infection in clinical patients. Methods The isolation of GBS from clinical specimens from an obstetrics and gynecology hospital in Shijiazhuang City between January 1, 2014 and December 31, 2017 was retrospectively analyzed, clinical distribution characteristics of GBS were analyzed, detection and antimicrobial resistance rates of GBS in different years were compared. Results A total of 2 368 strains of GBS were isolated, the main specimens were vaginal and anal sphincter swabs (2 229 strains, 94.13%), obstetric clinics and wards were the main detected wards (94.97%). A total of 2 229 strains of GBS were isolated from vaginal and anal sphincter swabs of pregnant women at 35 to 37 weeks of gestation, with a isolation rate of 3.68%. Isolation rate of GBS in vaginal and anal sphincter swabs of pregnant women during perinatal period was increasing year by year, difference was statistically significant (χ_trend²=44.78,P<0.05). Isolation rate of GBS was the highest in vaginal and anal sphincter swabs of pregnant women aged 25-34 years (3.69%). 2 368 strains of GBS had the highest resistance rate to azithromycin (78.21%), followed by erythromycin (77.62%), clindamycin (77.15%) and levofloxacin (47.68%), but all were sensitive to penicillin, vancomycin, linezolid, cefuroxime and ceftriaxone. Trend chi-square test results showed that resistance rates of GBS to erythromycin, azithromycin, clindamycin and levofloxacin increased year by year, with statistical significance (all P<0.05). Conclusion GBS isolated from pregnant women at 35-37 weeks of gestation in this hospital as well as antimicrobial resistance rate of GBS is increasing year by year, penicillin can be used as the preferred antimicrobial agent for treatment of GBS infection, clinicians should pay attention to the detection of GBS and the changing trend of antimicrobial resistance.