Kimon Chatzistamatiou

Primary Screening for Cervical Cancer Based on High-Risk Human Papillomavirus (HPV) Detection and HPV 16 and HPV 18 Genotyping, in Comparison to Cytology

Objectives The objective of the present study is to assess the performance of a high-risk human papillomavirus (HR-HPV) DNA test with individual HPV-16/HPV-18 genotyping as a method for primary cervical cancer screening compared with liquid-based cytology (LBC) in a population of Greek women taking part in routine cervical cancer screening. Methods The study, conducted by the “HEllenic Real life Multicentric cErvical Screening” (HERMES) study group, involved the recruitment of 4,009 women, aged 25–55, who took part in routine cervical screening at nine Gynecology Departments in Greece. At first visit cervical specimens were collected for LBC and HPV testing using the Roche Cobas 4800 system. Women found positive for either cytology or HPV were referred for colposcopy, whereas women negative for both tests will be retested after three years. The study is ongoing and the results of the first screening round are reported herein. Results Valid results for cytology and HPV testing were obtained for 3,993 women. The overall prevalence of HR-HPV was 12.7%, of HPV-16 2.7% and of HPV-18 1.4%. Of those referred for colposcopy, cervical intraepithelial neoplasia grade 2 or worse (CIN2+) was detected in 41 women (1.07%). At the threshold of CIN2+, cytology [atypical squamous cells of undetermined significance (ASC-US) or worse] and HPV testing showed a sensitivity of 53.7% and 100% respectively, without change between age groups. Cytology and HPV testing showed specificity of 96.8% and 90.3% respectively, which was increased in older women (≥30) in comparison to younger ones (25–29). Genotyping for HPV16/18 had similar accuracy to cytology for the detection of CIN2+ (sensitivity: 58.5%; specificity 97.5%) as well as for triage to colposcopy (sensitivity: 58.5% vs 53.7% for cytology). Conclusion HPV testing has much better sensitivity than cytology to identify high-grade cervical lesions with slightly lower specificity. HPV testing with individual HPV-16/HPV-18 genotyping could represent a more accurate methodology for primary cervical cancer screening in comparison to liquid-based cytology, especially in older women.

Epidemiology of HPV infection and current status of cervical cancer prevention in Greece: final results of the LYSISTRATA cross-sectional study.

The objective of this study was to assess the overall prevalence of the human papilloma virus (HPV) infection and distribution of high-risk HPV (hrHPV) types in Greece and evaluate the participation of women in primary and secondary cervical cancer prevention. This was a prospective, cross-sectional study carried out between October 2005 and January 2011 in Greece; 5379 women filled out the study questionnaire anonymously. 5107 women underwent cervical HPV-DNA testing, either by Hybrid Capture 2, followed by restriction fragment length polymorphism-PCR, or by the Abbott Real-Time High-Risk HPV test. Overall, 5.8% (295/5107) of women were positive for hrHPV infection. The most common hrHPV type was HPV-16 (24.8% among infected women; 1.4% overall), followed by HPV types 31, 35, 53, 18, 51, 56, 58, 52, 39, 66, 45, 33, 59, and 68. In respect to primary prevention of cervical cancer, acceptance of anti-HPV vaccination appeared to decrease over time (from 85–89.9% annually during 2005–2008 to 64.4–60.5% during 2009–2010, P<0.001). In respect to secondary prevention, only 30.3% of women had regular (annually for more than 5 years) Pap smears; regular gynecologic examinations, Papanicolaou testing, and knowledge of HPV were all associated with various demographic parameters (age, education, place of residence, occupation, and income). The prevalence of hrHPV infection in Greece is similar to that in other European countries; the most common type is HPV-16. The initially relatively high acceptance of HPV vaccination decreased after licensing of the vaccine. Demographic parameters appear to influence participation in cervical cancer screening.

Effect of mode of delivery on vertical human papillomavirus transmission - A meta-analysis.

A systematic review of the literature has been conducted (last update March 2014) for clinical studies reporting the prevalence of human papillomavirus (HPV) in the offspring of HPV-infected women in association to their mode of delivery. A meta-analysis was carried out according to the identification of concordant neonatal to maternal HPV types. Overall eight studies were included in the meta-analysis. Our pooled results, showed that caesarean section is associated with significantly lower rates of HPV transmission than vaginal birth (14.9% vs. 28.2%, risk ratio or RR: 0.515, 95% confidence interval or CI: 0.34–0.78). The number of caesarean sections needed to prevent one case of perinatal infection (number needed to treat or NNT) would be 7.5. As a conclusion it should be noted that caesarean section decreases the risk for perinatal HPV transmission by approximately 46%. Perinatal transmission still occurs in approximately 15% of the children born by caesarean section.

Diagnostic accuracy of high-risk HPV DNA genotyping for primary cervical cancer screening and triage of HPV-positive women, compared to cytology: preliminary results of the PIPAVIR study

PurposeThe purpose of the presented PIPAVIR (persistent infections with human papillomaviruses; http://www.pipavir.com) subanalysis is to assess the performance of high-risk (hr) HPV-DNA genotyping as a method of primary cervical cancer screening and triage of HPV positive women to colposcopy compared to liquid-based cytology (LBC) in an urban female population.MethodsWomen, aged 30–60, provided cervicovaginal samples at the Family-Planning Centre, Hippokratio Hospital of Thessaloniki, Greece, and the Department of Gynecology and Obstetrics in Mare Klinikum, Kiel, Germany. Cytology and HPV genotyping was performed using LBC and HPV Multiplex Genotyping (MPG), respectively. Women positive for cytology [atypical squamous cells of undetermined significance (ASC-US) or worse] or hrHPV were referred for colposcopy.ResultsAmong 1723/1762 women included in the final analysis, hrHPV and HPV16/18 prevalence was 17.7 and 9.6%, respectively. Cytology was ASCUS or worse in 7.6%. Cervical Intraepithelial Neoplasia grade 2 or worse (CIN2+) was detected in 28 women (1.6%). Sensitivity of cytology (ASCUS or worse) and HPV DNA testing for the detection of CIN2+ was 50.0 and 100%, and specificity was 94.49 and 85.49%, respectively. The screening approach according to which only women positive for HPV16/18 and for hrHPV(non16/18) with ASCUS or worse were referred to colposcopy presented 78.57% sensitivity and 13.17% positive predictive value (PPV).ConclusionsHPV testing represents a more sensitive methodology for primary cervical cancer screening compared to cytology. For triage of HPV positive women to colposcopy, partial HPV genotyping offers better sensitivity than cytology, at the cost of higher number of colposcopies.

Human papillomavirus E7 protein detection as a method of triage to colposcopy of HPV positive women, in comparison to genotyping and cytology. Final results of the PIPAVIR study

The objective of the presented cross‐sectional‐evaluation‐screening study is the clinical evaluation of high‐risk(hr)HPVE7‐protein detection as a triage method to colposcopy for hrHPV‐positive women, using a newly developed sandwich‐ELISA‐assay. Between 2013‐2015, 2424 women, 30‐60 years old, were recruited at the Hippokratio Hospital, Thessaloniki/Greece and the Im Mare Klinikum, Kiel/Germany, and provided a cervical sample used for Liquid Based Cytology, HPV DNA genotyping, and E7 detection using five different E7‐assays: “recomWell HPV16/18/45KJhigh”, “recomWell HPV16/18/45KJlow”, “recomWell HPV39/51/56/59”, “recomWell HPV16/31/33/35/52/58” and “recomWell HPVHRscreen” (for 16,18,31,33,35,39,45,51,52,56,58,59 E7), corresponding to different combinations of hrHPVE7‐proteins. Among 1473 women with eligible samples, those positive for cytology (ASCUS+ 7.2%), and/or hrHPV DNA (19.1%) were referred for colposcopy. Cervical Intraepithelial Neoplasia grade 2 or worse (CIN2+) was detected in 27 women (1.8%). For HPV16/18‐positive women with no triage, sensitivity, positive predictive value (PPV) and the number of colposcopies needed to detect one case of CIN2+ were 100.0%, 11.11% and 9.0 respectively. The respective values for E7‐testing as a triage method to colposcopy ranged from 75.0‐100.0%, 16.86‐26.08% and 3.83‐5.93. Sensitivity and PPV for cytology as triage for hrHPV(non16/18)‐positive women were 45.45% and 27.77%; for E7 test the respective values ranged from 72.72‐100.0% and 16.32‐25.0%. Triage of HPV 16/18‐positive women to colposcopy with the E7 test presents better performance than no triage, decreasing the number of colposcopies needed to detect one CIN2+. In addition, triage of hrHPV(non16/18)‐positive women with E7 test presents better sensitivity and slightly worse PPV than cytology, a fact that advocates for a full molecular screening approach.

Fetal growth impairment after first-trimester chorionic villus sampling: a case–control study

Abstract Objective: The objective of this study is to test if chorionic villus sampling (CVS) is associated with fetal growth impairment, after controlling for maternal and fetal factors. Study design: Case–control study of singleton fetuses whose mothers had undergone CVS (N = 442) and 2969 controls. The primary outcomes were the prevalence of birthweight < 10th centile and birthweight <3rd centile; the prevalence of preeclampsia was the secondary outcome. Tested predictors in logistic regression analysis included CVS, free beta-hCG MoMs, PAPP-A MoMs, first-trimester mean uterine artery pulsatility index (PI) z-scores, maternal height, BMI, age, and smoking. Results: The proportion of newborns with birthweight <10th centile (7.9 versus 6.2%), and <3rd centile (1.6 versus 1.1%) did not differ between the two groups. Maternal age, smoking during pregnancy, PAPP-A MoMs, and mean first-trimester uterine PI z-score were significant predictors for these outcomes. Although the prevalence of preeclampsia was higher in the CVS group (3.2 versus 1.3%, OR 2.62, 95% CI 1.41–4.89), the association was abolished in the regression analysis, in which maternal body mass index, free b-hCG levels, and mean first-trimester uterine PI z-score were the only significant predictors. Conclusions: CVS is not associated with fetal growth impairment, possibly because the resulting mechanical disruption is compensated by the developing placenta, without significantly impairing its function.

Cigarette Smoking Promotes Infection of Cervical Cells by High-Risk Human Papillomaviruses, but not Subsequent E7 Oncoprotein Expression

Persistent cervical infection with high-risk human papillomaviruses (hrHPVs) is a necessary, but not sufficient, condition for the development of cervical cancer. Therefore, there are other co-factors facilitating the hrHPV carcinogenic process, one of which is smoking. To assess the effect of smoking on high-risk (hr) HPV DNA positivity and on the expression of HPV E7 oncoprotein, as a surrogate of persistent hrHPV infection, we used data from women recruited for the PIPAVIR project, which examined the role of E7 protein detection in cervical cancer screening. Women were tested for hrHPV DNA, using Multiplex Genotyping (MPG), and E7 protein, using a novel sandwich ELISA method, and gave information on their smoking habits. Among 1473 women, hrHPV prevalence was 19.1%. The odds ratio (OR) for hrHPV positivity of smokers compared to non-smokers was 1.785 (95% confidence intervals (CI): 1.365–2.332, p < 0.001). The ORs for E7 positivity, concerning hrHPV positive women, ranged from 0.720 to 1.360 depending on the E7 detection assay used, but this was not statistically significant. Smoking increases the probability of hrHPV infection, and smoking intensity is positively associated to this increase. Smoking is not related to an increased probability of E7 protein positivity for hrHPV positive women.

Self-collected cervicovaginal sampling for site-of-care primary HPV-based cervical cancer screening: a pilot study in a rural underserved Greek population

Abstract In the present pilot study, the feasibility of a site-of-care cervicovaginal self-sampling methodology for HPV-based screening was tested in 346 women residing in underserved rural areas of Northern Greece. These women provided self-collected cervicovaginal sample along with a study questionnaire. Following molecular testing, using the cobas® HPV Test, Roche®, HPV positive women, were referred to colposcopy and upon abnormal findings, to biopsy and treatment. Participation rate was 100%. Regular pap-test examination was reported for 17.1%. Among hrHPV testing, 11.9% were positive and colposcopy/biopsy revealed 2 CIN3 cases. Non-compliance was the most prevalent reason for no previous attendance. Most women reported non-difficulty and non-discomfort in self-sampling (77.6% and 82.4%, respectively). They would choose self-sampling over clinician-sampling (86.2%), and should self-sampling being available, they would test themselves more regularly (92.3%). In conclusion, self-sampling is feasible and well-accepted for HPV-based screening, and could increase population coverage in underserved areas, helping towards successful prevention.

A New Sandwich ELISA Test Simultaneously Detecting E7 Proteins of HPV-16,18 and 45 in Cervical Smears

Persistent infections by high-risk papilloma viruses (HPV) are the main etiological factor for cervical cancer, and E7 oncoproteins were suggested as new markers for tumor progression. The objective of this study was to generate a new Enzyme-Linked Immunosorbent Assay (ELISA)-based detection system to monitor expression of the E7 proteins of the high-risk HPV (hrHPV) types HPV-16, HPV-18, and HPV-45 in cervical smears. Using a combination of rabbit monoclonal antibodies raised against E7 proteins of HPV-16 and HPV-18/HPV-45, respectively, a trivalent E7-ELISA was developed and validated, using recombinant E7 proteins of various HPV types and lysates from E7- positive cervical carcinoma cells. The amount of 0.5 picogram of E7 protein per well was determined as detection limit. The E7-ELISA was used to determine E7 protein levels in cervical smears obtained from a total of 67 women. E7 protein concentration was below the detection limit in all HPV-negative smears, and E7 protein concentrations above background were found in some HPV-positive cervical samples. Together the work described herein provides a new tool for the simultaneous detection of E7 proteins of the three most prevalent in cervical neoplasia hrHPV types.

Serum relaxin and cervical length for the prediction of spontaneous preterm birth in second‐trimester symptomatic women

To investigate whether serum relaxin level is associated with preterm birth in symptomatic women, either as a standalone test or in the context of a combined model of serum relaxin and cervical length (CL).