Fig. 1

Histological findings and HPV detection. a Apocrine acrosyringeal keratosis associated with syringocystoadenoma papilliferum. Higher magnification of a warty-like area of apocrine acrosyringeal keratosis (b) and of a cystic area of syringocystoadenoma papilliferum (c). The arrow in the inset indicates a cell with perinuclear halo suggestive of a possible HPV infection. d RCA. Genomic DNA extracted and purified from the paraffin embedded sample was amplified with TempliPhi Amplification Kit (Amersham Biosciences, Milan, Italy) according to the manufacturer's instruction except an higher (450 mM) concentration of nucleotides. To resolve the concatemers, RCA products were digested with BamHI, (Invitrogen-Life Technologies, Monza, Italy), for 3 h at 37 °C in a total volume of 20 μl. Digestion products were resolved by 0.8 % agarose gel electrophoresis. W12 is a cell line containing episomal HPV16. M is a Xlarge DNA Ladder (GeneDirex,Rome, Italy). The arrow indicates the amplified band of about 8000 bp. e RT-PCR. Total RNA extracted and purified from the paraffin embedded sample was subjected to retro-transcription and nested PCR with degenerate primers. Amplified products were resolved in ethidium bromide stained agarose gel. W12 is a cell line expressing HPV16 mRNA. No RT is a control in which reverse transcriptase was omitted to exclude the presence of contaminating genomic DNA. M5 is a DNA Molecular Weight Marker V (Roche, Milan, Italy). The arrow indicates the amplified products. f ISH: In situ hybridization was performed with ZytoFast kit (Bioptica, Milan, Italy). Specific probes for HPV 89 were prepared by the asymmetric PCR [16] with consensus primers. The arrows indicate cells with HPV positive green/blue stained nuclei. Since consecutive sections were not available, the corresponding localization in H/E preparation cannot be displayed