Alterations in human papillomavirus-related biomarkers after treatment of cervical intraepithelial neoplasia G. Valasoulis a, ⁎, G. Koliopoulos a , C. Founta a , M. Kyrgiou b , I. Tsoumpou c , O. Valari a , P. Martin-Hirsch d , A. Daponte e , P. Karakitsos f , E. Paraskevaidis a a University Hospital of Ioannina, Ioannina, Greece b Queen Charlotte's and Chelsea Hospital, Hammersmith Hospital, London, UK c University Hospitals of South Manchester, Manchester, UK d Central Lancashire Teaching Hospitals, Preston, UK e University Hospital of Larissa, Larissa, Greece f Attikon University Hospital, Athens, Greece abstract article info Article history: Received 1 September 2010 Available online 8 January 2011 Keywords: Cervical intraepithelial neoplasia (CIN) Loop electrosurgical excision procedure (LEEP) Human papillomavirus (HPV) Biomarkers HPV DNA-typing HPV mRNA p16 NASBA Flow cytometry Objective. This study aims to assess the alterations in various HPV-related biomarkers 6 months post- treatment and how these relate to various risk factors and individual characteristics; their role for the prediction of treatment failure was also evaluated. Material and methods. Design: Prospective observational study. Population: Women planning to undergo treatment for cervical intraepithelial neoplasia. Intervention: A liquid-based cytology sample was taken pre-operatively. This was tested for HPV genotyping, Nucleic Acid Sequence Based Amplification, flow cytometric evaluation and p16 immunostaining. A repeat LBC sample was obtained 6 months post-treatment and was tested for the same biomarkers. Outcomes: The alterations of the biomarkers 6 months post-treatment were recorded. Their relation to individual characteristics and risk factors (age, smoking, sexual history, use of condom, CIN grade, excision margin status, crypt involvement) as well as their role for the prediction of residual/recurrent disease were assessed. Analysis: The accuracy parameters (sensitivity, specificity, positive and negative predictive value and the likelihood ratios) of each biomarker for the prediction of recurrent/residual CIN were calculated. Results. A total of 190 women were recruited. All biomarkers had significantly higher negativity rates post- treatment compared to pre-treatment ones. Multivariate analysis demonstrated that consistent condom use post- treatment significantly reduces the high-risk HPV positivity rates in comparison to no use (OR=0.18; 95% CI: 0.09–0.38). Sensitivity and specificity for all high risk HPV DNA testing were 0.5/0.62, respectively; the relevant values for only type 16 or 18 DNA typing were 0.5/0.92, for NASBA 0.5/0.94, for flow 0.5/0.85 and for p16 0.25/0.93. Conclusion. CIN treatment reduces positivity for all HPV-related biomarkers. Consistent condom use significantly reduces high-risk HPV positivity rates. More cases of treatment failures are required in order to specify whether different combinations of HPV-related biomarkers could enhance the accuracy of follow up, possibly in the form of a Scoring System that could allow tailored post-treatment surveillance. © 2010 Elsevier Inc. All rights reserved. Introduction Cervical Intraepithelial Neoplasia (CIN) is the result of persistent infection with high-risk Human Papillomavirus (HPV) types [1]. It is well established that in general, cervical cancer precursors can be effectively treated by conservative methods, either destructive or excisional. Most of the available treatment modalities present similar and high efficacy in eradicating intraepithelial lesions [2–5], but they can also, inevitably, have failures. The reported failure rates range considerably between different centers and vary between 5 and 30% depending on a variety of factors such as marginal status of the tissue excised, patient's age and smoking with the pooled rate being reported at 10% [6–10]. As a result of treatment failures there is a reported increased risk (4–5 times) for invasive cervical cancer in women who have been treated for CIN compared to the general population [2,11–13]. It is imperative to identify women with persistent or recurrent disease so that they can be retreated and thus reduce the risk for future invasive disease development. The follow up for women who have undergone treatment for CIN traditionally consists of regular Pap smears with or without colpo- scopy in certain circumstances or centers [14]. In the last decade HPV DNA testing, without typing, was advocated for use in post-treatment Gynecologic Oncology 121 (2011) 43–48 ⁎ Corresponding author. Research Fellow in Obstetrics and Gynaecology, Neochor- opoulo Ioannina, PO BOX 253, 45500, Ioannina, Greece. Fax: +30 2651099224. E-mail address: gvalasoulis@gmail.com (G. Valasoulis). 0090-8258/$ – see front matter © 2010 Elsevier Inc. All rights reserved. doi:10.1016/j.ygyno.2010.12.003 Contents lists available at ScienceDirect Gynecologic Oncology journal homepage: www.elsevier.com/locate/ygyno