POLYMERASE CHAIN-REACTION ANALYSIS OF HUMAN PAPILLOMAVIRUS IN ARCHIVAL CERVICAL CYTOLOGIC SMEARS

We used a nested polymerase chain reaction (PCR) to study the presence of human papillomavirus (HPV) DNA in stained, archival cervical cytol ogic smears, where MYO9/MY11 served as outer primers and GP5/GP6 and a s inner primers. It was found to give a higher positivity rate than PC R using the EI degenerate consensus primers, where the sensitivity was decreased to 80%. It seemed optimal to use less sample DNA (0.5%) for the reaction; larger volumes resulted in decreased reactivity. Simila rly, the presence of bovine serum albumin helped to improve the reacti vity. The risk of cross-contamination did not seem to be a major obsta cle to a valid analysis. The prevalence of HPV in normal smears was 10 %, and in the high grade squamous intraepithelial lesion group it was 80%. Smears with cytologic evidence of HPV gave 100% positivity, while those containing cancer cells gave 80%. In patients whose Southern bl ot had demonstrated the presence of HPV, 87% of the simultaneously tak en smears were also positive with nested PCR. Similarly, in those whos e Southern blot analysis was negative, the corresponding smear was pos itive in 41%; this reactivity Tons associated with simultaneous squamo us intraepithelial lesions. The prevalence values indicated that this analysis is both sensitive and specific and that it can be used to eva luate the performance of other diagnostic methods. The validity is suf ficient to allow retrospective cohort studies of the natural history o f HPV infections during carcinogenesis.