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Diagnostic performance of p16/Ki-67 dual immunostaining at different number of positive cells in cervical smears in women referred for colposcopy

Ursula Salobir Gajsek
Ursula Salobir Gajsek
, 
Andraz Dovnik
Andraz Dovnik
, 
Iztok Takac
Iztok Takac
, 
Urska Ivanus
Urska Ivanus
, 
Tine Jerman
Tine Jerman
, 
Simona Sramek Zatler
Simona Sramek Zatler
 y 
Alenka Repse Fokter
Alenka Repse Fokter
   | 19 nov 2021
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Article Category: Research Article
Publicado en línea: 19 nov 2021
Páginas: 426 - 432
Recibido: 07 jul 2020
Aceptado: 28 sept 2021
DOI: https://doi.org/10.2478/raon-2021-0043
© 2021 Ursula Salobir Gajsek, Andraz Dovnik, Iztok Takac, Urska Ivanus, Tine Jerman, Simona Sramek Zatler, Alenka Repse Fokter, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Figure 1

Positive reaction was defined as p16 brown signal and Ki-67 red signal (red arrow) present in the same cell with red stained nucleus and brown stained cytoplasm. Note: negative p16/Ki-67 dual immunostaining (p16/Ki-67 DS) reaction (black arrow) (p16/Ki-76 DS, magnification 400x).
Positive reaction was defined as p16 brown signal and Ki-67 red signal (red arrow) present in the same cell with red stained nucleus and brown stained cytoplasm. Note: negative p16/Ki-67 dual immunostaining (p16/Ki-67 DS) reaction (black arrow) (p16/Ki-76 DS, magnification 400x).

Figure 2

The association between the number of p16/Ki-67 dual immunostaining (p16/Ki-67 DS) positive cells and the risk for cervical intraepithelial neoplasia (CIN2+). Observed values are marked as points. Smoothed line (Method spline) is added for better trend representation.
The association between the number of p16/Ki-67 dual immunostaining (p16/Ki-67 DS) positive cells and the risk for cervical intraepithelial neoplasia (CIN2+). Observed values are marked as points. Smoothed line (Method spline) is added for better trend representation.

Figure 3

Diagnostic performance of p16/Ki-67 dual immunostaining (p16/Ki-67 DS) at different cut-offs (number of positive cells).
Diagnostic performance of p16/Ki-67 dual immunostaining (p16/Ki-67 DS) at different cut-offs (number of positive cells).

Cervical intraepithelial neoplasia (CIN2+) according to the number of p16/Ki-67 dual immunostaining (p16/Ki-67 DS) positive cells

p16/Ki-67 Histology
Positive cells n < CIN2 n (%) CIN2+ n (%)
0 91 86 (94.5) 5 (5.5)
1 17 15 (88.2) 2 (11.8)
2 10 10 (100.0) 0 (0.0)
3 4 3 (75.0) 1 (25.0)
4 3 2 (66.7) 1 (33.3)
≥ 5 49 16 (32.7) 33 (67.3)
Total 174 132 (75.9) 42 (24.1)

Diagnostic performance of p16/Ki-67 dual immunostaining (p16/Ki-67 DS) according to cytology results and according to different cut-offs (number of positive cells) in detecting cervical intraepithelial neoplasia (CIN2+)

CIN2+(n) p16/Ki-67
Sensitivity (%, 95% CI) Specificity (%, 95% CI) PPV† (%, 95% CI) NPV‡ (%, 95% CI)
Cytology result
Negative 66.7 70.4 6.5 98.6
(n = 101) 3 (9.4–99.2) (60.3–79.2) (0.8–21.4) (92.3–100.0)
ASC-US 6 50.0 61.1 30.0 78.6
(n = 24) (11.8–88.2) (35.7–82.7) (6.7–65.2) (49.2–95.3)
LSIL 2 50.0 50.0 14.3 85.7
(n = 14) (1.3–98.7) (21.1–78.9) (0.4–57.9) (42.1–99.6)
HSIL 100.0 0.0 89.7
(n = 29) 26 (86.8–100.0) (0.0–70.8) (72.6–97.8) /
Number of positive p16/Ki-67 cells cut-off
1+ 88.1 65.2 44.6 94.5
(n = 174) 42 (74.4–96.0) (56.4–73.2) (33.7–55.9) (87.6–98.2)
2+ 42 83.3 76.5 53.0 93.5
(n = 174) (68.6–93.0) (68.4–83.5) (40.3–65.4) (87.1–97.4)
3+ 42 83.3 84.1 62.5 94.1
(n = 174) (68.6–93.0) (76.7–89.9) (48.5–75.1) (88.2–97.6)
4+ 42 81.0 86.4 65.4 93.4
(n = 174) (65.9–91.4) (79.3–91.7) (50.9–78.0) (87.5–97.1)
5+ 78.6 87.9 67.3 92.8
(n = 174) 42 (63.2–89.7) (81.1–92.9) (52.5–80.1) (86.8–96.7)

p16/Ki-67 dual immunostaining (p16/Ki-67 DS) positivity and number of positive cells among different smear results

Cervical cytology Number of p16/Ki-67 positive cells (n, [%])
 Total
0 (Negative) 1 2 3 4 ≥ 5 1+ (Total Positive)
Normal 70 (69.3) 13 (12.9) 9 (8.9) 1 (1.0) 1 (1.0) 7 (6.9) 31 (30.7) 101
ASC-US 14 (58.3) 0 (0.0) 1 (4.2) 2 (8.3) 1 (4.2) 6 (25.0) 10 (41.7) 24
LSIL 7 (50.0) 3 (21.4) 0 (0.0) 0 (0.0) 1 (7.1) 3 (21.4) 7 (50.0) 14
AGC-N 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 1 (100.0) 1 (100.0) 1
HSIL 0 (0.0) 1 (3.4) 0 (0.0) 0 (0.0) 0 (0.0) 28 (96.6) 29 (100.0) 29
ASC-H 0 (0.0) 0 (0.0) 0 (0.0) 1(25.0) 0 (0.0) 3 (75.0) 4 (100.0) 4
Inv. cancer 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 0 (0.0) 1(100.0) 1 (100.0) 1
Total 91 (52.3) 17 (9.8) 10 (5.7) 4 (2.3) 3 (1.7) 49 (28.2) 83 (47.7) 174
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