p16/Ki67 dual stain triage versus cytology in primary human papillomavirus‐based cervical cancer screening with limited genotyping

Abstract

The introduction of primary human papillomavirus (HPV) cervical cancer screening requires the implementation of an appropriate triage strategy that will be effective in detecting high‐grade cervical disease without losing diagnostic specificity. From the 30.066 screening tests results, a total of 1086 with available high‐risk human papillomavirus (HRHPV) with limited genotyping, cytology, and p16/Ki67 dual‐stain were selected. Two triage strategies for primary HPV screening were analyzed retrospectively based on the study group. Performance characteristics for p16/Ki67 and cytology triage in the detection of cervical intraepithelial neoplasia grade 2 or worse (CIN2+) and grade 3 or worse (CIN3+) were calculated, detected in colposcopic biopsy. In HPV16/18‐positive cases, primary HPV with p16/Ki67 triage was significantly more specific than cytology (53.1%/16.8% for CIN2+; p < 0.0001; 45.9%/17.0% for CIN3+; p < 0.0001), with yielded sensitivity (95.7%/84.8% for CIN2+; p = 0.0955; 100.0%/87.5% for CIN3+; p = 0.0832). In other HRHPV‐positive cases (N16/N18), p16/Ki67 triage was also significantly higher specific (51.3%/15.3% for CIN2+; p < 0.0001; 44.5%/16.5% for CIN3+; p < 0.0001), with sensitivity (92.3%/74.4% for CIN2+; p = 0.0522; 90.9%/81.8% for CIN3+; p = 0.5637). Diagnostic predictive values were significantly higher for p16/Ki67 triage with the highest PPV in HPV16/18‐positive cases for CIN2+ (45.4%; 95% confidence interval [CI]: 35.2–55.8; p < 0.0001) and very high NPV in all HPV‐positive cases regardless of detected genotype (96.3%–100.0%). The risk (1‐NPV) for CIN3+ in HRHPV16/18‐positive/p16/Ki67‐negative women was 0.0%. Superior diagnostic performance compared to cytology for detecting cervical cancer precursors indicates that p16/Ki67 dual‐immunostain may be a highly effective tool of triage in primary HPV screening with limited HPV 16/18 genotyping in secondary cervical cancer prevention.

Should we use risk selection tests for HPV 16 and/or 18 positive cases: Comparison of p16/Ki67 and cytology

“…A retrospective analysis of cytologic‐virologic‐immunocytochemical‐histologic test results was conducted, and the PPV of cytology and DS as triage tests for specific major cervical cancer screening abnormalities was evaluated. The study data were retrieved from the Center's registry, with a detailed methodology previously described in a series of related studies 33–36 …”

“…According to a manufacturer's manual, a DS‐positive test result was defined as at least one observed epithelial cell with simultaneous red nuclear stain for Ki67 and brown cytoplasmatic stain for p16 in the same epithelial cell 37 . A detailed protocol for an evaluation of immunoexpression was presented previously 33–36 …”

“…The study data were retrieved from the Center's registry, with a detailed methodology previously described in a series of related studies. [33][34][35][36] Major screening abnormalities were defined in this study as screening abnormalities with potentially higher HSIL/CIN2+ risk that justifies a direct referral to colposcopy of HPV 16/18+ patients without prior triage testing, identified through primary HPV-based screening with limited genotyping. Management in these cases was according to ASCCP 2019 Risk-Based Management Consensus Guidelines.…”

“…Detection of HR-HPV was performed using the Abbott RealTime High Risk HPV molecular in vitro PCR test (Abbott Molecular), in adherence to the manufacturer's instructions. This test phenotypes 12 types of highrisk HPV DNA (31,33,35,39,45,51,52,56,58,59, 66, and 68), as well as genotypes HPV 16 and 18 (limited genotyping). Test results were classified as HPV 16/18+ if either of these types was detected, and as HPV HR12+ if one or more of the other 12 non-16 and non-18 HR-HPV types were detected.…”

“…[38][39][40][41] The requirements of minimal colposcopy protocol included endocervical sampling (curettage and brushing), direct biopsies for all abnormal colposcopic findings or those suggestive for invasion, or additional random biopsies as per Polish protocols. All histologic diagnoses of cervical biopsies and endocervical sampling were reviewed by a gynecological pathologist according to the LAST 2012/WHO 2014 terminology42,43 with more details available in a series of related studies 33. Triage models analyzed Three HSIL/CIN2+ risk triage models for HPV 16/18+ cases in primary HPV-based cervical cancer screening were compared and analyzed.…”