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.