p16/Ki-67 dual staining in cervico-vaginal cytology: Correlation with histology, Human Papillomavirus detection and genotyping in women undergoing colposcopy

STI Unit, San Gallicano Dermatological Institute, via Elio Chianesi 53, 00144, Rome, Italy.
Gynecologic Oncology (Impact Factor: 3.77). 05/2012; 126(2):198-202. DOI: 10.1016/j.ygyno.2012.05.004
Source: PubMed


To evaluate the CINtec PLUS assay (mtm laboratories), a new immunocytochemical method for the simultaneous detection of p16(INK4a) and Ki-67, in liquid-based cervico-vaginal cytology, investigating the association of the dual staining with HPV infection and genotyping as well as cytological and histological abnormalities.
140 women with a cervico-vaginal sample obtained immediately before the colposcopy were enrolled. This cytological sample was used for HPV testing with the Linear Array HPV Genotyping Test, the dual staining with the CINtec PLUS kit and the morphology assessment.
Cytology results were 38 NILM, 16 ASC-US, 32L-SIL, 54H-SIL or worse. 113 patients also had a colposcopy-guided biopsy, classified as 14 negative, 35 CIN1, 24 CIN2, 37 CIN3, 3 invasive SCC. A strong association between p16/Ki-67 and HR-HPV infection was found (COR=6.86, 95% CI: 1.84-31.14). Importantly, the association between p16/Ki-67 positivity and HPV16 and/or 18 infection was 2-fold stronger compared to that with the infection by other HR-HPV types (COR=9.92, 95% CI: 2.39-47.77 vs COR=4.20, 95% CI: 0.99-20.87). In addition, p16/Ki-67 positivity rate significantly increased with the severity of the cytological and histological abnormalities (p<0.05 in both cases). p16/Ki-67 positivity resulted strongly associated with a CIN2+ diagnosis (COR=10.86 95% CI: 4.16-29.12).
This preliminary study evidenced that p16/Ki-67 immunostaining might have a relevant clinical role, since the dual staining was significantly associated with HR-HPV infection, particularly with HPV 16 and 18, and the increasing grade of the cervical lesions, the positivity for this biomarker being strongly related to the presence of a CIN2+ lesion.

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    • "Recently, a study in Germany with women of over thirty years old demonstrated a new screening model for HPV using the marking for the p16 and ki-67 proteins simultaneously, the efficiency of this type of screening was verified as an important factor in the progression of viral infection and cancer pathogenesis (Petry et al., 2011). Donà et al. (2012) analyzed cervical smears from 140 women, the samples went through ki-67 and p16 quantification process simultaneously via CINtec® PLUS, the results evidenced in this study corroborate previous data, showing that the immunohistochemistry for these markers may have an important diagnostic value, and the positivity for these biomarkers is highly related to the presence of high-grade premalignant lesions. Other immunohistochemistry kits with simultaneous action for p16 and Ki-67 have been proven effective in the early diagnosis of cervical cancer, the use of ProEx™ immunohistochemistry demonstrated results with sufficiently effective accuracy, and also highlighting the need for repetition in only one third of the samples (Walts et al., 2009). "
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    ABSTRACT: he critical sensitivity and specificity of the screening system for detection of HPV and cervical cancer indicates a necessity to study new biomarkers of early viral infection. This study aimed to review t he literature evidence of novel biomarkers of viral progression and cervical cancer. To conduct this review, the electronic databases Medline, SciELO, Pubmed, in Portuguese and English, referring to biomarkers of viral progression and HPV pathogenesis were consulted . The viral infectious and carcinogenic action of HPV among the biomarkers of tumor progression described in the literature was observed, with highlights to microRNAs (miRNA) Transglutaminase Type 2 (TG2), Ki - 67, p16 INK4a and Dynamin2 and HPV L1 capsid Protein . Based on scientific evidence, it was concluded that it is possible to improve the process by feasible techniques of high predictive value, thus highlighting the need for further studies to prove the biomarker effectiveness for subsequent us e in the national screening system.
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    ABSTRACT: Objective: This two-arm longitudinal study was performed within a regional organized cervical-cancer-prevention program in which HPV-DNA test is used in primary screening. The aim was to analyze the diagnostic performances of p16INK4a/Ki-67 dual-test and E6/E7-mRNA test in identifying CIN2+ lesion among HPV-DNA positive (HPV-DNAve) women triaged for LSIL-or-worse liquid based cytology (LBC). Methods: Thirty-six thousand thirty-one women participated to HPV-DNA screening program pilot study. Three thousand six hundred forty-one resulted HPV-DNAve; among these, 43% were LSIL-or-worse (LSIL+). HPV-DNAve/LSIL+ patients were submitted to colposcopy and histological assessment of any visible lesions. Dual-test was performed on 794 residual LBC specimens. In 405 cases, dual-test result was related to histology, considering CIN2+ as endpoint. mRNA test has been carried out retrospectively, on a subset of 173 residual LBC specimens. Results: Agreement between dual-test and histological diagnosis was 59%. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of cytology-plus-dual-test approach were 62.3%, 76.8%, 63.1% and 84.2%, respectively. Dual-test improved specificity, PPV and NPV of cytological triage Agreement between mRNA testing and histology was 65%. Cytology-plus mRNA testing showing sensitivity, specificity, PPV and NPV reaching 32.1%, 94.9%, 75% and 50%, respectively; implemented specificity and PPV of cytology alone in triaging DNA-ve/LSIL+ patients (p<0.01). Conclusions: We provided promising data indicating the important role that p16(INK4)/Ki-67 dual-test, and mostly E6/E7 mRNA test, might have in triaging HPV-DNAve. These approaches would exclude the occurrence of cervical cancer and would avoid overtreatment, at the same time. Further longitudinal analysis has to be considered.
    Gynecologic Oncology 11/2012; 128(3). DOI:10.1016/j.ygyno.2012.11.030 · 3.77 Impact Factor
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    ABSTRACT: Pap test, and especially HPV DNA test, identify a large group of women who do not have any clinically relevant lesions, i.e., CIN2+ (Cervical Intraepithelial Neoplasia grade 2 or worse), but who are at greater risk of getting lesions in the future. The follow up of these women needs new biomarkers with prognostic value. The objective of this study is to evaluate the prognostic value of E6/E7 mRNA over-expression assay (PreTect HPV-Proofer, Norchip) for 5 HR-HPV types (16, 18, 31, 33, and 45) for progression to CIN2+ after a negative colposcopy. This prospective study, conducted at four Italian centres, enrolled 673 women with either a negative colposcopy or a negative or CIN1 histology. The clinical end-point was histological confirmation of CIN2+. Women were classified at baseline according to mRNA results and managed according to local colposcopy protocols. At least one conclusive follow-up test was obtained for 347 women (25 months average lapse since recruitment, range 5-74). Only seven CIN2+ were detected during follow up, three among the 82 women positive for mRNA at baseline, two among the 250 negative (Fisher exact test, p = 0.02), and two among the 12 with an invalid test. Absolute CIN2+ risk was 6.7/1,000 person/years in the whole cohort. The absolute CIN2+ risk was 18.4/1,000 person/years and 3.6/1,000 person/years in mRNA-positive and mRNA-negative women, respectively. In conclusion, E6/E7 mRNA over-expression appears to be a good candidate as a prognostic biomarker to manage HR-HPV DNA-positive women with negative colposcopy or histology, particularly in order to decrease follow-up intensity in those who are negative.
    PLoS ONE 02/2013; 8(2):e57600. DOI:10.1371/journal.pone.0057600 · 3.23 Impact Factor
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