Detection of human papillomavirus DNA in cell scrapes and formalin-fixed, paraffin-embedded tissue of the uterine cervix by filter in situ hybridisation.
Filter in situ hybridisation (FISH) was used to detect the presence of DNA of human papillomavirus (HPV) types 6/11 or 16/18 in cell scrapes (CYTOFISH) and formalin-fixed, paraffin-embedded biopsies (HISTOFISH) taken from the uterine cervices of 19 women. Paraffin tissue sections collected for HISTOFISH were either digested with pepsin or lysed with alkali/Triton X-100. The digest or lysate of the tissue sections and cell scrapes were applied to nylon or nitrocellulose membranes for nucleic acid hybridisation using 32P-labeled HPV-DNA probes. CYTOFISH and HISTOFISH were compared directly by taking samples for each method from the cervices of the same women. Of 19 women examined by colposcopy, cytology, and histology, eight were assessed as normal and 11 had evidence of a cervical disorder and/or the presence of HPV infection. Whereas no HPV-DNA was detected in the normal cases, the presence of HPV-DNAs was detected by both CYTOFISH and HISTOFISH in 11 cases with histological evidence of HPV infection and/or dysplasia. In these HPV positive cases, eight contained HPV 16/18, two HPV 6/11, and one a mixed infection of HPV 6/11/16/18. The high correlation between the results of CYTOFISH and HISTOFISH shows that formalin-fixed, paraffin-embedded cervical biopsies are suitable specimens for the detection and typing of HPV-DNA by FISH. Both CYTOFISH and HISTOFISH should facilitate studies on the prevalence and distribution of HPVs and their association with neoplasia.
Demeter
T
T
Department of Microbiology, University of Western Australia, Nedlands.
Kulski
J K
JK
Rakoczy
P
P
Sterrett
G F
GF
Pixley
E C
EC
D001345:Autoradiography; D001706:Biopsy; D002460:Cell Line; D015344:DNA Probes, HPV; D004279:DNA, Viral / Q000032:analysis*; D005260:Female; D005557:Formaldehyde; D006801:Humans; D015151:Immunoblotting; D027383:Papillomaviridae / Q000302:isolation & purification; D010232:Paraffin; D012680:Sensitivity and Specificity; D014412:Tumor Virus Infections / Q000382:microbiology* / Q000473:pathology; D002577:Uterine Cervical Diseases / Q000382:microbiology* / Q000473:pathology
D015344:DNA Probes, HPV; D004279:DNA, Viral; D005557:Formaldehyde; D010232:Paraffin