A retrospective study was conducted to reveal the natural history of cervical carcinoma infected with human papillomavirus (HPV) and to document latent persistence of HPV infection. Thirty-eight formalin-fixed, paraffin-embedded hysterectomy specimens of cervical carcinoma and cervical intraepithelial neoplasia (CIN) III were examined for the presence of HPV 6, 11, 16, 18, 31, 33, and 35 DNA by in situ hybridization (ISH) with biotinylated DNA probes. The HPV 6 and 11 were not detected, but HPV 16, 18, 31, 33, and 35 were detected in 63% (24 of 38) of CIN III and cervical carcinoma. The HPV-negative specimens by ISH were subjected to polymerase chain reaction (PCR) for detection of HPV 16 and 52b with high sensitivity. In 11 of 14 specimens, specific amplification of HPV 16 was detected and HPV DNA was demonstrated in 92% (35 of 38). A retrospective examination for the presence of HPV DNA by ISH and PCR was performed on sequential biopsy specimens of eight cases whose hysterectomy specimens were judged as HPV 16/18 positive. In the eight cases, HPV 16 was constantly demonstrated in all biopsy specimens throughout the course, from early dysplastic change to carcinoma. This finding indicates long (up to 10 years) persistence of HPV infection in the natural course of progression to carcinoma of the uterine cervix.