Purpose: The purpose of this study was to determine the prevalence of human papillomavirus (HPV) in the population in Kunming, Yunnan, China, before and after the introduction of HPV preventive vaccines. Materials and Methods: In total, 28,959 patients were enrolled in this study between 1 January 2016 and 31 August 2019. HPVs were genotyped using a flow-through hybridisation technique, and differences in HPV infection rates before and after the introduction of an HPV vaccine were determined. Results: The prevalence of HPV before and after the introduction of HPV vaccines was 17.74% and 17.11%, respectively. This difference was not statistically significant (χ2 = 1.920, P > 0.05). The HPV infection rates showed a bimodal U-shaped curve for all age groups. The most common genotypes of HPV detected were HPV52, HPV16, HPV58 and HPV39. Conclusion: Although the overall HPV infection rate in Kunming did not change significantly after the introduction of HPV vaccines, differences in HPV infection rates and multi-typic HPV infection rates were evident in certain age groups.