Sensitive detection and typing of human papillomavirus DNA in gynecological cell scrapings by slot-blot hybridization.
A rapid and sensitive method for detecting and typing human papillomaviruses (HPVs) in cell scrapings is presented. DNA from scrapings is extracted and bound to nitrocellulose filters (Slot-Blot). By DNA-DNA hybridization with specific 32P-labelled HPV-probes (types 6/11 or 16/18) the patient's DNA is then analyzed for the presence of, and for the type of, HPV DNA sequences. A parallel hybridization with a human repetitive element (Alu sequence) allows quantitation of the different hybridization results. Experiments with HeLa cell DNA show that as little as 10(4) HPV sequences can be detected and typed specifically with this test. Evaluation of this test is completed within 6 to 7 days after cell collection. This Slot-Blot method was used to analyse 1330 specimens taken at the Bernese Dysplasia Outpatient Clinic. The results reveal a very high percentage (90%) of HPV-positive cases in the patient group examined.
Gerber-Huber
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Institut für Biochemie und Molekularbiologie, Universität Bern, Switzerland.
Gorgievski-Hrisoho
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Meandzija
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Locher
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GW
D001345:Autoradiography; D015139:Blotting, Southern; D015344:DNA Probes, HPV; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D005836:Genitalia, Female / Q000032:analysis*; D006367:HeLa Cells; D006801:Humans; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000032:analysis*; D010759:Phosphorus Isotopes; D002578:Uterine Cervical Dysplasia / Q000175:diagnosis*
D015344:DNA Probes, HPV; D004279:DNA, Viral; D010759:Phosphorus Isotopes