# High-Risk HPV CISH Detection in Cervical Biopsies with Weak and/or Focal p16 Immunohistochemical Positivity

## TL;DR

This study shows that cervical biopsies with weak or focal p16 positivity can still contain high-risk HPV DNA, suggesting the need for closer monitoring and additional testing.

## Contribution

The study demonstrates the presence of high-risk HPV DNA in cervical biopsies with weak/focal p16 positivity using CISH, which was previously considered nonspecific.

## Key findings

- HPV DNA was detected in 60% of cases with weak/focal p16 positivity using CISH.
- Different nuclear positivity patterns were observed, including punctate, diffuse, and mixed.
- The findings suggest that weak/focal p16 positivity may still indicate HPV infection, especially with increased Ki67.

## Abstract

In cervical biopsies, for diagnosis of Human Papilloma Virus (HPV) related conditions, the immunohistochemical staining for p16 has a diagnostic value only if diffusely and strongly positive, pattern named “block-like”. “Weak and/or focal (w/f) p16 expression” is commonly considered nonspecific. In our previous study, we demonstrated the presence of high-risk HPV (hrHPV) DNA by LiPa method in biopsies showing w/f p16 positivity. The aim of the present study was to investigate the presence of hrHPV-DNA by CISH in the areas showing w/f p16 expression. We assessed the presence of hrHPV16, 18, 31, 33, 51 by CISH in a group of 20 cervical biopsies showing w/f p16 expression, some with increased Ki67, and in 10 cases of block-like expression, employed as control. The immunohistochemical p16 expression was also assessed by digital pathology. hrHPV-CISH nuclear positivity was encountered in 12/20 cases of w/f p16 expression (60%). Different patterns of nuclear positivity were identified, classified as punctate, diffuse and mixed, with different epithelial distributions. Our results, albeit in a limited casuistry, show the presence of HPV in an integrated status highlighted by CISH in w/f p16 positive cases. This could suggest the necessity of a careful follow-up of the patients with “weak” and/or “focal” immunohistochemical patterns of p16, mainly in cases of increased Ki67 cell proliferation index, supplemented with molecular biology examinations.