Triage of LSIL/ASC-US with p16/Ki-67 dual staining and human papillomavirus testing: a 2-year prospective study

Abstract

Objective: To investigate human papillomavirus (HPV) DNA testing and p16/Ki-67 staining for detecting cervical intraepithelial grade 2 or worse (CIN2+) and CIN3 in women referred to colposcopy with minor abnormal cervical cytology low-grade squamous intraepithelial lesions (LSIL) and atypical squamous cells of undermined significance (ASC-US). The clinical performance of both tests was evaluated as stand-alone tests and combined, for detection CIN2+ and CIN3 over 2 years. Methods: ThinPrep â liquid-based cytology (LBC) specimens were collected from 1349 women with repeat LSIL or ASC-US. HPV DNA was performed using Hybrid Capture. Where adequate material remained (n = 471), p16/Ki-67 overexpression was assessed. Clinical performance for detection of histologically diagnosed CIN2+ and CIN3 was calculated.Results: Approximately 62.2% of the population were positive for HPV DNA, and 30.4% were positive for p16/Ki-67. p16/Ki-67 showed no significant difference in positivity between LSIL and ASC-US referrals (34.3% versus 28.6%; P = 0.189). Women under 30 years had a higher rate of p16/Ki-67 compared to those over 30 years (36.0% versus 26.6%; P = 0.029). Overall HPV DNA testing produced a high sensitivity for detection of CIN3 of 95.8% compared to 79.2% for p16/Ki-67. In contrast, p16/Ki-67 expression offered a higher specificity, 75.2% versus 40.4% for detection of CIN3. Combining p16/Ki-67 with HPV DNA improved the accuracy in distinguishing between CIN3 and <CIN3. The absolute risk of CIN3 increased from 15.6% in women who were HPV DNA positive to 27% in women positive for HPV DNA and p16/Ki-67. Those negative for HPV DNA and p16/Ki-67 had a low risk of 1.2% of CIN3. Conclusion: The addition of p16/Ki-67 to HPV DNA testing leads to a more accurate stratification of CIN in women presenting with minor cytological abnormalities.

Dual staining for p16/Ki‐67 to detect high‐grade cervical lesions: Results from the Screening Triage Ascertaining Intraepithelial Neoplasia by Immunostain Testing study

“…populations. A recent meta-analysis 6 of studies published between 2012 and 2017 demonstrated that dual-stained cytology was more specific than high-risk HPV testing for the identification of CIN2+; the relative specificity was 1.65 (95% CI: 1.42-1.92, P < .0001, n = 11 studies 8,14,15,[18][19][20][21][22][23][24][25] for the triage of ASC-US and 2.45 (95% CI:…”

Meta‐analysis of the accuracy of p16 or p16/Ki‐67 immunocytochemistry versus HPV testing for the detection of CIN2+/CIN3+ in triage of women with minor abnormal cytology

“…Twenty studies reported the accuracy of p16, among which 1 study also evaluated dual staining. Eighteen studies evaluated the accuracy of dual staining only. The positivity criteria for p16 staining varied across the studies.…”

“…Eight studies used the nuclear scoring system of Wentzensen et al For 1 study, the p16 positivity cutoff was brown cytoplasmic staining for p16 in more than 10 cervical cells. With respect to dual staining, 19 studies considered simultaneous red nuclear and brown cytoplasmic staining of at least 1 cervical cell as a positivity criterion.…”

Triage of high-risk HPV positive women in cervical cancer screening

“…In triage of hrHPV-positive women with a negative cytology result, sensitivity for ≥CIN2 varies between 67% and 92%, with a specificity of 73-82% [36,37]. In triage of women with hrHPV-positive low-grade cytology, sensitivity and specificity of ≥CIN2 vary from 75-94% to 51-88%, respectively [38][39][40][41][42][43]. Limited studies have been performed with immediate p16/Ki-67 triage of hrHPV-positive women, without additional cytological interference.…”