p16 INK4a immunocytochemistry in liquid - based cervicovaginal specimens with modified Papanicolaou counterstaining.

2006 
Aim: To evaluate the feasibility and usefulness of a modified Papanicolaou counterstain for p16 INK4a immunostaining in liquid-based cervicovaginal samples. Methods: Immunocytochemistry with p16 INK4a and modified Papanicolaou counterstain was performed on 81 liquid-based samples including 23 cases within normal limits (WNL), 6 with low-grade squamous intraepithelial lesion (LSIL), 20 with high-grade squamous intraepithelial lesion (HSIL), 16 with atypical squamous cells of undetermined significance (ASC-US) and 16 with atypical squamous cells, highgrade lesion cannot be excluded (ASC-H). Results were compared with histological or cytological follow-up. For comparison, 29 further cases (10 LSIL, 10 ASC-H and 9 HSIL) were immunostained with p16 INK4a and conventionally counterstained with hematoxylin. Intensity of immunostain in SIL cases was assessed using a 0 to 3 scoring system. Interobserver agreement was calculated by Kappa statistics. Results: p16 INK4a expression was detected in 13% of WNL, 66.6% of LSIL, 100% of HSIL, 31.3% of ASC-US and 81.3% of ASC-H. Excluding two cases with no residual dysplastic cells in the immunocytochemistry, all cases with CIN2-3 at follow-up expressed p16 INK4a and none of the p16 INK4a negative cases showed a high-grade lesion at follow-up. No evident differences in pattern or intensity of p16 INK4a expression between study and control group were observed. Interobserver agreement was significantly better in the study group compared with the conventional immunostain (combined Kappa value 0.773 vs. 0.549, p<0.05), and still
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