Human papillomavirus (HPV) infection is associated with multiple types of cancer, but the evidence has not yet been fully elucidated in bladder cancer. Frozen tissue samples collected from 146 patients aged 32 to 89 years-old with bladder cancer pathological diagnosis between 2015 and 2019 were analyzed. HPV genotyping and integration status determination were performed by capture-based next generation sequencing. Statistical analysis of HPV type distributions was performed according to stage, grade, gender and age group of patients. Mean age of 146 patients was 66.64 ± 10.06 years and 83.56% were male. Overall HPV infection rate was 28.77% (37.5% in female and 27% in male), with 11.90% HPV integration events. Among them, 17.12% single and 11.65% co-infections were observed. HPV18 (24.66%) was the most prevalent genotype, followed by HPV33, 16, 39. All HPV were European lineage (A). HPV16 was more prevalent in female (p=0.04). HPV infection may contribute a causative role both in men and women with bladder cancer. HPV18, followed by HPV 33, 16 and 39 genotypes potentially represent the predominant oncogenic risk types of bladder carcinogenesis. © The Author(s) 2020. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.