Use of the same archival Papanicolaou smears for detection of human papillomavirus by cytology and polymerase chain reaction

An optimal method for the processing of archival cervical Papanicolaou (pap )-stained smears for the amplification of human papillomavirus (HPV) DNA by polymerase chain reaction (PCR) was developed. This methodology was then a pplied to a series of 44 pap smears designated as HPV positive or negative (on the basis of both major and minor cytological criteria) or cervical int raepithelial neoplasia (CIN)-cancer. For the detection of HPV DNA, each sam ple was tested with the consensus GP5/6 primers, and when negative, with CP I-IIG primers. The HPV DNA was detected in 100% (8 of 8) of GIN-cancer smea rs using the GP5/6 primers. In smears with cytological evidence of HPV with out CMI the use of both sets of primers yielded positive results in 100% (1 9 of 19) of the samples. Direct sequence analysis of PCR products showed th at 16 of the 27 HPV-positive samples contained more recently described HPV types. When tested with both primer combinations, all 17 cytologically nega tive smears were positive for beta-globin but negative for HPV DNA. The fin dings show the value of using archival pap smears for further investigation s to address issues such as latency, but they indicate that cytological cri teria and DNA technology will be critical factors in the reliability of the results.