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

Abstract

We compared clinical performance of p16/Ki-67 dual-stained cytology and human papillomavirus (HPV) genotyping, via different algorithms-alone, or in combination with cytology-to identify cervical intraepithelial neoplasia grade 2 or worse (CIN2+) and grade 3 or worse (CIN3+) in women referred to as colposcopy. We included 492 cervical specimens (134 normal, 130 CIN1, 99 CIN2, 121 CIN3, 8 cancers) randomly selected from 1158 specimens with valid conventional cytology, HPV (cobas 4800 HPV test) and biopsy results. Dual-stained cytology was retrospectively performed (CINtec PLUS assay) on PreservCyt material; slides were read by a cytologist and confirmed by two pathologists, blinded to cytology, biopsy and genotyping results. Sensitivity and specificity (95% confidence intervals in parentheses) of dual-stained cytology to detect CIN2+ and CIN3+ were compared to other screening tests available for the same women. Positivity rate for dual-stained cytology increased with histological severity: 30.6% in normal, 41.5% in CIN1, 72.7% in CIN2, 86.8% in CIN3 and 87.5% in cancer. Dual-stained cytology alone had lower sensitivity than HPV testing for CIN2+ [80.7% (75.0-85.6) vs 89.9% (85.3-93.5)] and CIN3+ [86.8% (79.7-92.1) vs 92.3% (86.2-96.2)]. However, corresponding specificity values were higher [64.0% (57.9-69.8) vs 56.1% (49.8-62.1) for CIN2+; 54.0% (48.7-59.2) vs 44.4% (39.2-49.6) for CIN3+]. Combining dual-stained cytology with an ASC-US abnormality threshold decreased specificity to 31.4% (25.9-37.4) for CIN2+ and 24.2% (19.9-29.0) for CIN3+. The corresponding values considering low squamous intraepithelial lesion threshold values were 42.8% (36.8-49.0) and 35.0% (30.1-40.1). Dual-stained cytology and HPV Abbreviations: AGC, atypical glandular cells; ASC-H, atypical squamous cells cannot exclude HSIL; ASC-US, atypical squamous cells of undetermined significance; CIN, cervical intraepithelial neoplasia; HPV, human papillomavirus; HSIL, high squamous intraepithelial lesion; LSIL, low squamous intraepithelial lesion; NILM, negative for intraepithelial lesion or malignancy; STAIN-IT, Screening Triage Ascertaining Intraepithelial Neoplasia by Immunostain Testing.

Comparison of CINtec PLUS cytology and cobas HPV test for triaging Canadian patients with LSIL cytology referred to colposcopy: A two-year prospective study

“…The significantly higher specificity of CINtec PLUS than HPV test was consistently observed, and in this respect, we note that CINtec PLUS is currently approved for triaging those testing positive in HPV primary screening [ 29 ]. The lower sensitivity, albeit with the caveats noted, and higher specificity of CINtec PLUS compared to HPV test we observed is consistent with a screening triage study recently reported in a Canadian population [ 23 ] and many other studies [ 17 , 21 , 30 , 31 ].…”

“…CINtec PLUS has been shown to be more sensitive than cytology with equal specificity, and more specific than HPV testing with relatively comparable sensitivity for detecting cervical intraepithelial neoplasia grade 2 or worse (CIN2 ) in LSILs [ 11 , 17 , 18 , 19 , 20 ]. While the clinical applications of CINtec PLUS in LSIL triage have been assessed in several studies in Europe and elsewhere [ 11 , 18 , 21 , 22 ], there has been limited evaluation of this method to serve as an adjunct test in LSIL triage in North American settings [ 23 ].…”

Novel biomarkers with promising benefits for diagnosis of cervical neoplasia: a systematic review

“…For miRNA (miR-9) assay, three studies recorded sensitivity, specificity and area under the ROC curve ranges of 52.9-67.3%, 76.4-94.4%%, and 0.71-0.85, respectively based on RT-qPCR platform using serum and tissue samples [2,11,53], with77.8% PPV and 70.2% NPV reported by Park et al [11]. 3) For the p16INK4a / ki-67 assays, twenty studies included reported sensitivity, specificity, PPV, NPV, and AUC ranges of 50-100%, 39-90.4%, 11.1-92.3%, 86.7-100%, and 0.76-0.90, respectively, based on immunocytochemistry (ICC) platform using liquid based cytology (LBC) samples [12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30] . On the other hand, four studies evaluating Vascular Endothelial Growth Factor (VEGF) assay reported sensitivity, specificity, and AUC ranges of 56-83.5%, 74.6-96%, and 0.83-0.88, respectively, based on Elisa platform using serum [5,[50][51][52], with Lawicki et al being the only author who reported PPV and NPV at 86 and 82%, respectively.…”

The Role of p16/Ki-67 Immunostaining, hTERC Amplification and Fibronectin in Predicting Cervical Cancer Progression: A Systematic Review

“…p16/Ki-67 IHC has been used most frequently throughout the studies. Similar to p16 and Ki-67 IHC alone, an increase in the number of DS-positive biopsy specimens was correlated with a more severe histological diagnosis [ 32 , 41 , 42 , 43 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 ] and with HPV DNA positivity [ 41 , 42 , 43 , 58 , 59 , 63 ] ( Table 2 ).…”

“…p16/Ki-67 IHC has been used most frequently throughout the studies. Similar to p16 and Ki-67 IHC alone, an increase in the number of DS-positive biopsy specimens was correlated with a more severe histological diagnosis [32,[41][42][43][53][54][55][56][57][58][59][60][61][62][63] and with HPV DNA positivity [41][42][43]58,59,63] (Table 2). Ki-67 was generally expressed in combination with p16 IHC, as DS positivity.…”