To understand human papillomavirus (HPV) and expression of osteopontin (OPN) in cervical diseased tissues, HPV infection was detected in paraffin-embedded specimens of cervical lesions from 90 patients with cervical cancer. Three polymerase chain reaction (PCR) techniques were used to determine the detectable rate of HPV infection. Expression of HPV OPN protein was detected using immunohistochemical methods. When a pairwise comparison was made among the three PCR methods, χ2 analysis indicated P>0.05 for detection through the methods of MY09/11 and GP5+/6+, and P>0.05 through the methods of MY09/11 and Nested-PCR. This indicated that there was no statistically significant difference in the HPV infection detection sensitivity of these methods. However, χ2 comparison of the methods of GP5+/6+ and Nested-PCR indicated P<0.05, which demonstrated that there was a statistically significant difference. The rate of positive HPV DNA measured with Nested-PCR was significantly higher than that measured using the GP5+/6+ PCR method. The HPV OPN protein is expressed in cervical cancer, and the HPV OPN polypeptide antibody has broad spectrum reaction capacity and significant multivalence for HPV infection. Immunohistochemical detection was performed on tissue specimens using the purified rabbit HPV OPN polypeptide antibody. Sixty-one cases exhibited a positive result and 29 a negative result. The total rate of positive detection was 67.78%. HPV OPN may therefore serve as a candidate target for tumor treatment, including targeted therapies and vaccine development.