A one-step method for detecting and typing human papillomavirus DNA in cervical scrape specimens from women with cervical dysplasia.
We studied 66 women with a previous dysplastic cervical cytological smear who were referred for colposcopy and biopsy for the presence of human papillomavirus (HPV) types 6, 11, 16, 18, and 31 DNA in cervical specimens. The specimens were analyzed by a novel hybridization method termed reverse blotting, in which cellular DNA is radiolabeled and used to probe a battery of cloned HPV DNAs immobilized on nitrocellulose. Reconstruction experiments demonstrated that this method could detect about one HPV genome equivalent per cell. HPV DNA sequences were detected in 52 (96%) of 54 patients who showed either condylomatous changes or dysplasia by cervical biopsy. HPV-16 was most commonly detected overall and was detected in 61% of moderate or severe dysplastic samples. HPV DNA was also detected in seven of 12 cervical scrapes from women with a history of dysplasia but with either normal or inflammatory changes noted on cervical biopsy. Our results indicate that the reverse-blot method can detect DNA homologous to various HPV types in a single experiment using DNA from the small numbers of cells obtained by cervical scraping.
Webb
D H
DH
Department of Medicine, Indiana University School of Medicine, Indianapolis.
Rogers
R E
RE
Fife
K H
KH
D002584:Cervix Uteri / Q000382:microbiology; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000032:analysis* / Q000302:isolation & purification; D005260:Female; D005814:Genes, Viral; D006801:Humans; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000145:classification / Q000235:genetics*; D014412:Tumor Virus Infections / Q000382:microbiology*; D002578:Uterine Cervical Dysplasia / Q000382:microbiology*
D004279:DNA, Viral; D004262:DNA Restriction Enzymes