In situ hybridization with digoxigenin-labeled DNA of human papillomaviruses (HPV 16/18) in HeLa and SiHa cells.
To establish a nonradioactive method for demonstrating HPV DNA in routinely treated smears of the uterine cervix (alcohol fixation, staining according to Papanicolaou, preservation), in situ hybridizations were carried out in HeLa and SiHa cells grown on slides. After detailed investigations, the sensitivity and specificity of the biotin-avidin method (10) initially used proved to be inadequate for this purpose. Demonstration of HPV 16 DNA in SiHa cells (SiHa cells only contain 1-2 HPV genome copies) was possible only by use of digoxigenin-labeled HPV 16 gene probes, as well as an improved purification of the sample DNA from vector contaminations. Thus, for the first time a protocol for correlation of the results of an in situ hybridization with the cytological appraisal in the very same smear preparation has been developed for routine diagnostics.
Heiles
H B
HB
Universität zu Köln.
Genersch
E
E
Kessler
C
C
Neumann
R
R
Eggers
H J
HJ
D001709:Biotechnology; D004279:DNA, Viral / Q000302:isolation & purification*; D004076:Digoxigenin; D005260:Female; D006367:HeLa Cells; D006801:Humans; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000302:isolation & purification*; D014407:Tumor Cells, Cultured / Q000382:microbiology; D002583:Uterine Cervical Neoplasms / Q000382:microbiology
D004279:DNA, Viral; D004076:Digoxigenin