Table 1.
Results of the HPV MLPA-Assay in Frozen Uterine Cervical Tissue Samples, Validated by qPCR, PapilloCheck, and FISH
| Diag. | Case | Age | % Cells | Typing |
Viral load |
Physical status |
Telomerase genes gain |
High risk class. MLPA | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| MLPA | qPCR | PapilloCheck | MLPA | qPCR* Ct HPV16/18 | qPCR† HPV16 | MLPA | qPCR† E2/E6 | MLPA TERT | MLPA TERC | |||||
| Normal no lesion (n = 7) | 1 | 64 | − | − | − | − | 35/37 | − | − | − | − | − | − | |
| 2‡ | 47 | − | − | − | − | 36/37 | − | − | − | − | − | − | ||
| 3 | 52 | − | − | − | − | 35/− | − | − | − | − | − | − | ||
| 4 | 52 | 16 | − | − | 0.11 | 36/− | − | epi | − | − | − | − | ||
| 5 | 73 | 16 | − | − | 0.14 | 35/40 | − | epi | − | − | − | − | ||
| 6 | 38 | 16 | 16 | 16 | 0.22 | 34/40 | − | epi | − | − | − | − | ||
| 7 | 60 | 16 | 16 | 16 | 2.10 | 34/38 | 0.16 | epi | int | − | − | − | ||
| Normal adj. to lesion (n = 20) | 8 | 75 | − | − | − | − | 38/− | n.d. | − | − | − | − | − | |
| 9 | 35 | − | 16 | − | − | 34/− | n.d. | − | − | − | − | − | ||
| 10 | − | − | − | − | − | 38/38 | n.d. | − | − | − | − | − | ||
| 11 | 47 | − | − | − | − | 35/− | n.d. | − | − | − | − | − | ||
| 12 | 36 | − | − | − | − | 37/− | n.d. | − | − | − | − | − | ||
| 13 | 43 | − | − | 45 | − | −/− | n.d. | − | − | − | − | − | ||
| 14 | 50 | 16 | − | 16 | 0.004 | 37/− | − | epi | − | − | − | − | ||
| 15 | 34 | 16 | 16 | − | 0.05 | 34/39 | 0.05 | epi | mix | − | − | − | ||
| 16 | − | 16 | 16 | 16 | 0.07 | 33/− | 0.02 | epi | mix | − | − | − | ||
| 17 | 32 | 16 | 16 | 58 | 0.13 | 32/− | 0.1 | epi | mix | − | − | − | ||
| 18 | − | 16 | 16 | 16; 59 | 0.18 | 34/− | 0.04 | epi | mix | − | − | − | ||
| 19 | 52 | 16 | 16; 31 | n.c. | 0.18 | 34/39 | 0.05 | epi | epi | − | − | − | ||
| 20 | − | 16 | 16 | 16 | 0.33 | 31/40 | 0.33 | mix (a)§ | mix | − | − | + | ||
| 21 | 64 | 16 | 16 | n.c. | 0.48 | 31/− | 0.22 | epi | mix | − | − | − | ||
| 22 | − | 16 | 16 | 16; 59 | 0.51 | 31/38 | 0.17 | epi | mix | − | − | − | ||
| 23 | 49 | 16 | 16 | 16 | 0.91 | 32/− | 0.34 | epi | n.c. | − | − | − | ||
| 24 | 32 | 16 | 16 | 16 | 2.5 | 30/− | 0.83 | epi | mix | − | − | − | ||
| 25 | 51 | 16 | 16 | n.c. | 6.8 | 26/38 | 1.6 | epi | epi | − | − | − | ||
| 26 | − | 16 | 16 | 16 | 13 | 26/− | 8 | epi | epi | − | − | − | ||
| 27 | − | 16 | 16 | 16 | 31 | 26/36 | 4.5 | epi¶ | mix | − | − | + | ||
| CIN1/condyloma (n = 10) | 28 | − | 60 | − | − | − | − | 37/− | n.d. | − | − | − | − | − |
| 29 | − | 60 | − | − | 52 | − | 39/40 | n.d. | − | − | − | − | − | |
| 30 | − | 95 | − | − | − | − | −/− | n.d. | − | − | − | − | − | |
| 31 | 22 | 40 | 16 | − | 56; 59 | 0.02 | 35/39 | − | epi | − | − | − | − | |
| 32 | − | 70 | 16 | 16 | 16 | 0.06 | 33/39 | − | epi | − | − | − | − | |
| 33 | 58 | 50 | 16 | 16 | 16; 59 | 0.09 | 33/37 | 0.09 | epi | epi | − | − | − | |
| 34 | 23 | 40 | 16 | 16 | 56; 59 | 0.14 | 33/39 | − | epi | − | − | − | − | |
| 35 | 32 | 90 | 16 | 16 | n.c. | 6.4 | 27/− | 2.4 | int (b)§ | int | − | − | + | |
| 36 | − | 60 | 16 | 16 | 16; 59 | 90 | 23/37 | 54 | epi | epi | − | − | + | |
| 37 | 21 | 90 | 16 | 16 | 16; 59 | 1008 | 19/− | 568 | epi¶ | epi | + | + | + | |
| CIN2 (n = 6) | 38 | 38 | 50 | 16 | 16; 45 | 16; 45 | 0.01 | 34/− | 0.03 | epi | mix | − | − | − |
| 39‡ | 43 | 60 | 16 | 16 | 16 | 1.4 | 29/38 | 0.76 | mix (c)¶ | mix | − | −¶ | + | |
| 40 | − | 70 | 18 | 18 | 16; 18; 59 | 1.7 | 35/27 | − | mix (b)§ | − | + | + | + | |
| 41 | − | 90 | 16 | 16 | 16; 33 | 50 | 23/− | 45 | epi | epi | + | + | + | |
| 42 | 59 | 70 | 16 | 16 | 16; 59 | 94 | 23/38 | 73 | epi | epi | + | + | + | |
| 43 | − | 60 | 16 | 16 | 16 | 130 | 22/− | 91 | epi§ | epi | − | − | + | |
| CIN3 (n = 7) | 44 | 53 | 50 | 16 | 16 | 16 | 0.65 | 31/38 | 0.14 | epi | mix | − | − | − |
| 45 | 30 | 70 | 16 | 16 | − | 3.1 | 28/− | 3.8 | int (b) | mix | − | − | + | |
| 46 | − | 30 | 16 | 16 | 16 | 5.9 | 27/40 | 15 | int (a) | int | − | + | + | |
| 47 | 49 | 50 | 16 | 16 | 16 | 8.4 | 27/− | 3.8 | epi | epi | − | − | − | |
| 48 | 36 | 50 | 16 | 16 | 16; 59 | 9.5 | 26/36 | 6.1 | mix (c) | epi | − | + | + | |
| 49 | − | 80 | 16 | 16 | 16 | 10 | 25/38 | 25 | epi | epi | − | + | + | |
| 50‡ | 27 | 60 | 16 | 16 | 16 | 249 | 24/− | 112 | epi¶ | epi | − | −¶ | + | |
| SCC (n = 17) | 51 | 30 | 95 | − | 45 | 45; 11 | − | 38/− | n.d. | − | n.d. | + | − | + |
| 52 | 44 | 30 | 16 | 33 | 16; 59; 33 | 0.01 | 35/− | − | epi | − | + | + | + | |
| 53 | − | 60 | 16 | 31 | 16 | 0.04 | 35/− | − | epi | − | − | − | − | |
| 54 | − | 80 | 18 | 18 | 16; 59 | 3.7 | 35/28 | − | epi§ | − | − | − | − | |
| 55 | 46 | 90 | 16 | 16 | − | 4.5 | 27/36 | 4.4 | int (b) | int | − | − | + | |
| 56 | − | 15 | 16 | 16 | 16 | 4.8 | 26/38 | 17 | int (b) | int | + | + | + | |
| 57 | 45 | 50 | 16 | 16 | 16 | 5.3 | 27/− | 12 | epi | n.c. | − | + | + | |
| 58 | − | 70 | 16 | 16 | 16 | 5.6 | 26/36 | 13 | epi | epi | − | + | + | |
| 59‡ | 51 | 80 | 16 | 16 | 16 | 9.4 | 27/40 | 3.7 | int (a) | int | − | − | + | |
| 60 | 70 | 95 | 16 | 16 | 16 | 25 | 24/− | 15 | epi | epi | − | − | + | |
| 61 | 50 | 60 | 16 | 16 | 16 | 62 | 23/39 | 81 | epi | epi | − | − | + | |
| 62 | 62 | 90 | 16 | 16 | 16 | 70 | 25/− | 23 | epi | epi | − | − | + | |
| 63 | 45 | 60 | 16 | 16 | 16 | 83 | 22/− | 153 | epi | epi | + | + | + | |
| 64 | 55 | 90 | 16 | 16 | 16 | 126 | 20/38 | 504 | mix (a) | mix | − | + | + | |
| 65 | 48 | 60 | 16 | 16 | 16; 59 | 134 | 22/37 | 38 | mix (b) | mix | − | + | + | |
| 66‡ | 58 | 80 | 16 | 16 | 16 | 149 | 20/37 | 198 | mix (b)¶ | mix | − | +¶ | + | |
| 67 | − | 70 | 16 | 16 | 16 | 166 | 20/37 | 404 | mix (a) | mix | − | + | + | |
% Cells, percentage (pre)malignant cells; HPV, human papillomavirus; MLPA, multiplex ligation-dependent probe amplification; qPCR, quantitative PCR; FISH, fluorescence in situ hybridization; +, detected; −, not detected; n.c., not conclusive; n.d., not determined; a, (partial) loss of target sequence 16E2.1; b, (partial) loss of target sequence 16/18E2.1 and (partial) loss of target sequence 16/18E2.2; c, (partial) loss of target sequence 16E2.2); epi, episomal; mix, episomal and integrated; int, integrated; TERC, telomerase RNA component; TERT, telomerase reverse transcriptase; high risk class; MLPA, high risk classification based on viral load of 25 HPV16 copies or more and/or viral integration and/or gain of the telomerase genes.
according to Lindh et al.23
according to Peitsaro et al.24
previously described by Theelen et al.6
FISH inconclusive.
Confirmed by FISH.