Evaluation of Biomarkers p16/Ki-67 in Cervical Cytology for Diagnosis of Cervical Intraepithelial Neoplasia

  • Shalini Rajaram
  • Sathija Puthiya kulap
  • Bindiya GuptaEmail author
  • V. K. Arora
  • Alok C. Bharti
  • Neerja Goel
Original Article



To test the clinical utility of biomarkers p16/Ki-67 expression in cervical cytology smears as a marker for transforming HPV infection.

Setting and Design

Experimental study at a tertiary care hospital.


Women who were screened positive on Pap and visual inspection tests (n = 280) underwent colposcopy and biopsy. p16/Ki-67 immunostaining was performed in abnormal Pap smears (n = 86), and HPV DNA testing was also performed in the same women.

Statistical Analysis

Sensitivity, specificity, negative predictive value, positive predictive value and likelihood ratios were calculated for each biomarker separately and in combination. McNemar test and receiver operating characteristic (ROC) curves were used to compare sensitivity and specificity of biomarkers with HPV DNA. Areas under the ROC curve (AUC) were compared using the Chi-square test.


Eighty-six women with abnormal cytology were evaluated with p16/Ki-67 immunocytochemistry; 20.9% (n = 18) and 18.6% (n = 16) were positive for each biomarker, while dual marker was positive in 15% (n = 13). In all smears, the sensitivity of p16INK4a/Ki-67 in detecting CIN 2+ lesion was 76.9% and specificity was 95.8%. For ASCUS (n = 42) and LSIL (n = 23) smears, specificity and negative predictive value of p16/Ki-67 for CIN 2+ were 100% with a likelihood ratio (LR+) of 27 and 25, respectively, suggesting good diagnostic accuracy. In comparison with HPV DNA testing, combined marker p16/Ki-67 was significantly more specific (p = 0.003); AUC was 0.734 and 0.635, respectively.


p16/Ki-67 evaluation in cervical cytology is a valuable biomarker in triaging for CIN 2+ disease in ASCUS and LSIL smears.


p16/Ki-67 HPV Cervical cancer screening Cervical cytology Biomarker 


Compliance with Ethical Standards

Conflict of interest

There is no conflict of interest among the authors.

Ethical Approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Informed Consent

Informed consent was obtained from all individual participants included in the study.


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Copyright information

© Association of Gynecologic Oncologists of India 2019

Authors and Affiliations

  1. 1.Department of Obstetrics and GynecologyUCMS & GTB HospitalDelhiIndia
  2. 2.Department of PathologyUniversity College of Medical SciencesDelhiIndia
  3. 3.Division of Molecular OncologyNational Institute of Cancer Prevention and Research NoidaUttar PradeshIndia
  4. 4.Molecular Oncology Laboratory, Department of ZoologyUniversity of DelhiDelhiIndia

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