Objective: To study the distribution of human papillomavirus (HPV) genotypes in population of hospital opportunistic screening and the relationship with cervical intraepithelial neoplasia (CIN) among women. Materials and Methods:A retrospective analysis was performed in 14,991 women who were undergoing HPV genotype testing and 253 inpatients who were undergoing conization pathological with CIN and with pre-HPV test at Jiangsu University Hospital between July 2013 and July 2015. Results: Of these, 25.99% were infected by HPV and 20.37% for high-risk HPV (hr-HPV), the highest prevalence of hr-HPV and multiple hr-HPV were all in the 55- to 64-year-old group (χ² = 37.125, p < 0.01, χ² = 43.668, p < 0.01). The most prevalent genotype is HPV52, then descending order were HPV16 and 58. On the basis of 253 inpatients, CIN I were 78, CIN II were 67, CIN III were 108. Before conization, hr-HPV positive in CIN I lesions were 74.36% (58/78) and CIN II were 94.03% (63/67), while in CIN III lesions there were 94.44% (102/108) (χ² =17.936, p < 0.01). The most frequent HPV genotype was HPV 16, followed by HPV 58 and 52. Infection with multiple- hr-HPV seemed to have no relationship with the severity of pathology increased (χ² =1.888, p > 0.01). Conclusion: This study established the prevalence of HPV genotypes that is similar to the population of CIN, except HPV 16 and HPV 52, 58 were also found in a large proportion of women, HPV 18 was not the most important. Therefore, in the future, a HPV vaccine covering HPV 52/16/58 should be used in this region compared to the currently available vaccine for the prevention strategy in the population.