Non-isotopic detection of HPV DNA in cervical smears using dot-blot hybridization.
Infection of the cervix with specific types of the human papillomavirus (HPV) results in condylomatous, dysplastic and/or neoplastic epithelial changes. To enable routine screening of premalignant cervical lesions for HPV DNA, we have developed a sensitive method of detecting HPV -6, -11, -16, -18 and -33 in cervical scrapes using a non-isotopic, dot-blot hybridization assay. Cloned, genomic HPV DNA is labelled with biotin-11-dUTP by nick translation, and used to screen for HPV types under high stringency conditions. DNA is extracted from cervical cells and spotted on nitrocellulose membranes. Cervical cells, from 20 patients attending colposcopy were screened for HPV types. All patients had a dysplasia or condyloma. HPV DNA was isolated from 80%. The method visualized an amount of target DNA as low as 1 pg without background. Compared with Southern blot hybridization utilizing radiolabelled probe, this assay is equally sensitive and specific. The technique is easy to do, and may significantly contribute to the management of cervical dysplasia.
Inoue
M
M
Department of Pathology, Foothills Hospital, Calgary, Alberta, Canada.
Duggan
M A
MA
Robertson
D I
DI
Chang-Poon
V
V
D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D027383:Papillomaviridae / Q000235:genetics*; D014626:Vaginal Smears
D004279:DNA, Viral