Objective To understand the pathogenic bacteria from children with vulvovaginitis and their antimicrobial resistance. Methods Clinical data of children with vulvovaginitis and positive bacterial culture of vaginal secretion from Beijing Children’s Hospital from January 1, 2016 to December 31, 2023 were analyzed retrospectively. Results Among 3 249 children with vulvovaginitis, a total of 3 389 pathogenic bacteria strains were detected, including 1 730 strains of Gram-negative bacteria and 1 659 strains of Gram-positive bacteria, accounting for 51.05% and 48.95%, respectively. The top 5 detected pathogens were Haemophilus influenzae (n=1 201, 35.44%), Streptococcus pyogenes (Group A) (n=694, 20.48%), Staphylococcus aureus (n=375, 11.06%), Streptococcus agalactiae (Group B) (n=183, 5.40%), and Haemophilus parainfluenzae (n=179, 5.28%). Among the isolated pathogens in pediatric patients across different age groups, Haemophilus influenzae accounted for 46.18% in children < 7 years old, Streptococcus pyogenes (Group A) and Haemophilus influenzae accounted for 27.08% and 25.86% respectively in children aged 7-11 years old, Streptococcus agalactiae (Group B) and Staphylococcus aureus accounted for 31.50% and 18.11% respectively in children >11-18 years old. The pathogens detected varied in different seasons. Among Gram-positive bacteria, Streptococcus pyogenes (Group A) had higher resistance rates to erythromycin and clindamycin, which were 93.52% and 92.94%, respectively. Among Gram-negative bacteria, Haemophilus influenzae had higher resistance rates to ampicillin, compound sulfamethoxazole, cefaclor, and cefuroxime (all >34% ). 648 Haemophilus influenzae strains (53.96%) and 40 Haemophilus parainfluenzae strains (22.35%) produced β-lactamase. Conclusion The common pathogenic bacteria causing infectious vulvovaginitis in children are Haemophilus influenzae, Streptococcus pyogenes (Group A), and Staphylococcus aureus. The detection rate of β-lactamase in Haemophilus influenzae is high. Understanding the pathogen detection and antimicrobial resistance status in hospitals can provide basis for clinician’s empirical and rational selection of antimicrobial agents.