Table 1.
Quantitative real-time PCR detection of HPV-16 and HPV-18 in LCM generated samples from squamous carcinomas of the cervix and in samples from adjacent normal epithelial and mesenchymal tissue. The first column lists the clinical/laboratory code of each of 26 cervical carcinomas. The second, third and fourth column list three numbers, in each case first the copy number of the virus per cell, then the CT value of the HPV-16/18 RT-PCR signal, and thirdly the CT value of the cellular marker, the HMBS gene. The second column lists data obtained with tumor samples, the third normal cell clusters flanking the tumors, and the fourth flanking mesenchymal tissues. Each CT value represents the average of the RT-PCR values obtained from two DNA preparations made from two juxtaposed LCM samples cleaved from the same thin section of the tumor. The abbreviations “pos.” and “neg.” refer to a positive or negative PCR signal, evaluated by gel electrophoresis, in cases where no quantitative real-time-PCR amplification could be achieved. The letter X replaces the copy number in those cases where the virus, but not the cellular gene as the denominator of the calculation could be quantified
| Name of tumor | HPV genome copy number (CT HPV/CT HMBS) | ||
|---|---|---|---|
| HPV-16 | tumor: | normal epithelium: | stroma: |
| T16–1 | 74 (14.8/20.0) | <1 (24.1/21.3) | <1 (27.9/26.6) |
| T16–2 | X (15.6/pos.) | 0 (neg./pos.) | 0 (neg./pos.) |
| T16–3 | X (14.5/pos.) | 0 (neg./pos.) | 0 (neg./pos.) |
| T16–4 | 630 (18.9/27.2) | 0 (neg./28.2) | 0 (neg./26.9) |
| T16–5 | 208 (14.5/21.8) | 2 (26.8/26.7) | <1 (27.0/24.0) |
| T16–6 | 104 (14.5/20.2) | <1 (29.3/22.6) | 0 (neg./22.3) |
| T16–7 | 9 (20.1/22.2) | <1 (25.1/21.5) | 0 (neg./24.0) |
| T16–8 | 49 (17.8/22.4) | <1 (32.0/22.3) | 0 (neg./25.3) |
| T16–9 | 16 (17.6/20.6) | <1 (24.2/21.7) | 0 (neg./22.3) |
| T16–10 | 97 (15.3/20.9) | <1 (28.0/23.4) | 0 (neg./25.0) |
| T16–11 | 128 (18.5/24.5) | <1 (29.8/26.3) | 0 (neg./27.1) |
| T16–12 | 181 (18.6/25.1) | 11 (24.6/27.1) | 0 (neg./27.7) |
| T16–13 | 315 (20.4/27.7) | 0 (neg./27.6) | 16 (24.6/27.6) |
| SiHa (600 cells) | 15 (24.9/28.8) | ||
| SiHa (60 cells) | 24 (28.3/32.9) | ||
| HPV-18 | |||
| T18–1 | 30 (18.3/22.2) | <1 (29.2/25.8) | 0 (neg./25.1) |
| T18–2 | 3 (22.0/22.5) | 0 (neg./23.0) | 0 (neg./25.1) |
| T18–3 | <1 (24.1/22.5) | <1 (26.5/25.0) | 0 (neg./23.7) |
| T18–4 | X (19.9/pos.) | X (30.3/pos.) | 0 (neg./pos.) |
| T18–5 | X (19.8/pos.) | 0 (neg./neg.) | 0 (neg./pos.) |
| T18–6 | X (15.6/n.d.) | X (26.3/neg.) | 0 (neg./neg.) |
| T18–7 | X (20.9/pos.) | X (29.2/neg.) | 0 (neg./neg.) |
| T18–8 | 2 (24.3/24.4) | 0 (neg./24.6) | 0 (neg./26.2) |
| T18–9 | 111 (19.0/24.8) | 0 (neg./27.7) | 0 (neg./27.4) |
| T18–10 | 6 (23.6/25.2) | 0 (neg./24.2) | 0 (neg./27.6) |
| T18–11 | 2 (21.5/21.5) | <1 (24.8/21.1) | <1 (26.5/23.3) |
| T18–12 | 1 (21.8/20.5) | <1 (28.5/22.5) | 0 (neg./26.4) |
| HeLa (600 cells) | 30 (25.8/30.6) | ||
| HeLa (60 cells) | 22 (29.1/34.3) | ||
| HPV-16/18: | |||
| T16/18–1 | 97 (18.3-HPV-18/23.9) | <1 (26.8-HPV-16/22.3) | 0 (neg./24.5) |