Sensitivity of digoxigenin and biotin labelled probes for detection of human papillomavirus by in situ hybridisation.
The sensitivity of digoxigenin and biotin labelled DNA probes for the detection of human papillomavirus (HPV) by dot blotting and in situ hybridisation was compared in tissues from cervical, laryngeal, and anogenital neoplasia. Probes were either labelled with digoxigenin by the random primer technique and detected with anti-digoxigenin antibody, or labelled with biotin by nick translation and detected with streptavidin, both methods having a common final visualisation procedure using alkaline phosphatase. Digoxigenin labelled probes proved two to 10-fold more sensitive by quantitative dot blotting and four-fold more sensitive in detecting HPV 16 DNA in a series of 31 anal carcinomas, compared with biotinylated probes. The digoxigenin method also produced less non-specific background staining of tissue sections than biotin labelled probes. It is concluded that digoxigenin DNA labelling and detection provides a simple, reliable, and efficient alternative to the use of biotin or radioactive isotopes for the detection of HPV DNA by in situ hybridisation. Digoxigenin labelled probes also offer the possibility of double labelling in situ hybridisation procedures when used with biotin labelled probes to provide simultaneous identification of different DNA sequences.
Morris
R G
RG
Department of Pathology, University Medical School, Edinburgh.
Arends
M J
MJ
Bishop
P E
PE
Sizer
K
K
Duvall
E
E
Bird
C C
CC
D001005:Anus Neoplasms / Q000382:microbiology; D001710:Biotin; D015139:Blotting, Southern; D015344:DNA Probes, HPV; D004076:Digoxigenin; D005260:Female; D005833:Genital Neoplasms, Female / Q000382:microbiology; D005834:Genital Neoplasms, Male / Q000382:microbiology; D006801:Humans; D007822:Laryngeal Neoplasms / Q000382:microbiology; D008297:Male; D009693:Nucleic Acid Hybridization; D027383:Papillomaviridae / Q000302:isolation & purification*; D014412:Tumor Virus Infections / Q000175:diagnosis*; D002583:Uterine Cervical Neoplasms / Q000382:microbiology
D015344:DNA Probes, HPV; D001710:Biotin; D004076:Digoxigenin