Abstract
Purpose
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.
Methods
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.
Results
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.
Conclusions
p16/Ki-67 evaluation in cervical cytology is a valuable biomarker in triaging for CIN 2+ disease in ASCUS and LSIL smears.
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References
World Health Organization (WHO): Cervical cancer estimated incidence, mortality and prevalence worldwide in 2018. http://www.iarc.fr/en/mediacentre/iarcnews/2018/gco_globocan2018.php. Accessed 15 Sept 2018.
Nanda K, McCrory DC, Myers ER, Bastian LA, Hasselblad V, Hickey JD, et al. Accuracy of the Papanicolaou test in screening for and follow-up of cervical cytologic abnormalities: a systematic review. Ann Intern Med. 2000;132(10):810–9.
Shastri SS, Dinshaw K, Amin G, Goswami S, Patil S, Chinoy R, et al. Concurrent evaluation of visual, cytological and HPV testing as screening methods for the early detection of cervical neoplasia in Mumbai, India. Bull World Health Organ. 2005;83(3):186–94.
Pourasad-Shahrak S, Salehi-Pourmehr H, Mostafa-Garebaghi P, Asghari-Jafarabadi M, Malakouti J, et al. Comparing the results of Pap smear and direct visual inspection (DVI) with 5% acetic acid in cervical cancer screening. Niger Med J. 2015;56:35–8.
Kulasingam SL, Hughes JP, Kiviat NB, Mao C, Weiss NS, Kuypers JM, et al. Evaluation of human papillomavirus testing in primary screening for cervical abnormalities: comparison of sensitivity, specificity, frequency of referral. JAMA. 2002;288:1749–57.
Petry KU, Schmidt D, Scherbring S, Luyten A, Reinecke-Lüthge A, Bergeron C, et al. Triaging Pap cytology negative, HPV positive cervical cancer screening results with p16/Ki-67 dual-stained cytology. Gynecol Oncol. 2011;121(3):505–9.
Schmidt D, Bergeron C, Denton KJ, Ridder R. p16/ki-67 dual-stain cytology in the triage of ASCUS and LSIL papanicolaou cytology: results from the European equivocal or mildly abnormal Papanicolaou cytology study. Cancer Cytopathol. 2011;119(3):158–66.
Kisser A, Zechmeister-Koss I. A systematic review of p16/Ki-67 immuno-testing for triage of low grade cervical cytology. BJOG. 2015;122(1):64–70.
A practical manual on visual screening for cervical neoplasia. Online screening material www.screening.iarc.fr. Accessed 15 Sept 2017.
Bornstein J, Bentley J, Bösze P, et al. 2011 colposcopic terminology of the international federation for cervical pathology and colposcopy. Obstet Gynecol. 2012;120:166–72.
Lapierre SG, Sauthier P, Mayrand M-H, Dufresne S, Petignat P, Provencher D, et al. Human papillomavirus (HPV) DNA triage of women with atypical squamous cells of undetermined significance with cobas 4800 HPV and hybrid capture 2 tests for detection of high-grade lesions of the uterine cervix. J Clin Microbiol. 2012;50(4):1240–4.
Possati-Resende JC, Fregnani JHTG, Kerr LM, Mauad EC, Longatto-Filho A, Scapulatempo-Neto C. The accuracy of p16/Ki-67 and HPV test in the detection of CIN2/3 in women diagnosed with ASC-US or LSIL. PLoS One. 2015;10(7):e0134445.
Cox JT, Schiffman M, Solomon D. Prospective follow-up suggests similar risk of subsequent cervical intraepithelial neoplasia grade 2 or 3 among women with cervical intraepithelial neoplasia grade 1 or negative colposcopy and directed biopsy. Am J Obstet Gynecol. 2003;188(6):1406–12.
Bibbo M, Klump W, DeCecco J. Procedure for immunocytochemical detection of p16INK4A antigen in thin-layer, liquid-based specimens. Acta Cytol. 2002;46:25–9.
Gustinucci D, Passamonti B, Cesarini E, Butera D, Palmieri EA, Bulletti S, et al. Role of p16(INK4a) cytology testing as an adjunct to enhance the diagnostic specificity and accuracy in human papillomavirus-positive women within an organized cervical cancer screening program. Acta Cytol. 2012;56:506–14.
Wang R, Li X, Qian M, Niu J, You Z. The natural history of cervical intraepithelial neoplasia I and the clinical significance of p16(INK4a) protein as a marker of progression in cervical intraepithelial neoplasia I. Zhonghua Fu Chan Ke Za Zhi. 2015;50(3):210–5.
Wentzensen N, Schwartz L, Zuna RE, et al. Performance of p16/Ki-67 immunostaining to detect cervical cancer precursors in a colposcopy referral population. Clin Cancer Res. 2012;18(15):4154–62.
Edgerton N, Cohen C, Siddiqui MT. Evaluation of CINtec PLUS testing as an adjunctive test in ASC- US diagnosed SurePath preparations. Diagn Cytopathol. 2013;41(1):35–40.
Bergeron C, Ikenberg H, Sideri M, Denton K, Bogers J, Schmidt D, PALMS Study Group, et al. Prospective evaluation of p16/Ki-67 dual-stained cytology for managing women with abnormal Papanicolaou cytology: PALMS study results. Cancer Cytopathol. 2015;123(6):373–81.
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This work was done at Department of Obstetrics and Gynecology and Department of Pathology, UCMS & GTB Hospital, Delhi, India.
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Rajaram, S., Puthiya kulap, S., Gupta, B. et al. Evaluation of Biomarkers p16/Ki-67 in Cervical Cytology for Diagnosis of Cervical Intraepithelial Neoplasia. Indian J Gynecol Oncolog 17, 46 (2019). https://doi.org/10.1007/s40944-019-0290-5
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DOI: https://doi.org/10.1007/s40944-019-0290-5