The present paper aims to investigate the relationship between human papillomavirus 16 (HPV16) infection in cervical diseases, and to provide scientific basis for prevention and treatment of cervical cancer. The proportion of HPV16 in different cervical diseases, the prevalence of HPV16 infection in the patients with cervical intraepithelial neoplasia (CIN) Ⅱ and higher diseases in different age groups, and the prevalence of simple and multiple infections of HPV were analyzed. A total of 1 057 patients infected with HPV were selected, and subjected to sub-typing for HPVs. The results revealed that 352 patients were HPV16 positive and the proportion of HPV16 detection was the highest in CIN Ⅲ group. The proportion of HPV16 positive was significantly different among varied cervical diseases (P<0.05) and it gradually increased along with the increased disease grade (P<0.05). There were significant differences in the prevalence of HPV16 infection in the patients with CIN Ⅱ and higher diseases among different age groups (P<0.05). The prevalence increased with the increased age (P<0.05). The prevalence of HPV16 infection in the patients with CIN Ⅱ and higher diseases was significant different between HPV simple and multiple infections (P<0.05). With types of HPV increasing, the prevalence showed a tendency of increase. The research indicates that there is certain relationship between HPV16 infection and high-grade cervical lesions. The older persons infected with HPV16 have the greater risks for cervical cancer. Taking the persistent infection of HPV16 into account, timely diagnosis and treatment for HPV infection could reduce the occurrence of high-grade CIN and cervical cancer.