Synthetic oligonucleotide probes for the detection of human papilloma viruses by in situ hybridisation

Abstract

Using the published nucleotide sequence data for human papilloma virus (HPV) types 1, 6 and 16, sequences of 30 bases in length from the beginning of the E6 open reading frame (ORF) were selected. Oligonucleotides were synthesised on an Oswel Gene Synthesizer and labelled at the 3' end with biotin using the enzyme terminal transferase. In situ hybridisation was carried out on paraffin sections of wart and cervical tissues mounted on silanated slides. A 2 h hybridisation step allowed the whole process to be completed within one working day. The technique successfully demonstrated the presence of HPV-1 in skin warts, and of HPV-6 and HPV-16 in genital warts and cervical lesions. This simple approach has diagnostic potential for the detection and typing of papilloma viruses in biopsy material.