The Role of HPV Genotyping, Cytology, and Methylation in the Triage of High-Risk HPV-Positive Patients

<b>Background/Objectives</b>: This study evaluates the diagnostic performance of DNA methylation testing, alone and in combination with cervical cytology, for high-grade squamous intraepithelial lesion (HSIL) detection. <b>Methods</b>: A prospective study was conducted on 170...

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Main Authors: Anastasia Mortaki, Athanasios Douligeris, Maria-Anastasia Daskalaki, Eleni-Sivylla Bikouvaraki, Eirini Louizou, George Daskalakis, Alexandros Rodolakis, Themos Grigoriadis, Kalliopi I Pappa
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Language:English
Published: MDPI AG 2025-05-01
Series:Biomedicines
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Online Access:https://www.mdpi.com/2227-9059/13/5/1139
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author Anastasia Mortaki
Athanasios Douligeris
Maria-Anastasia Daskalaki
Eleni-Sivylla Bikouvaraki
Eirini Louizou
George Daskalakis
Alexandros Rodolakis
Themos Grigoriadis
Kalliopi I Pappa
author_facet Anastasia Mortaki
Athanasios Douligeris
Maria-Anastasia Daskalaki
Eleni-Sivylla Bikouvaraki
Eirini Louizou
George Daskalakis
Alexandros Rodolakis
Themos Grigoriadis
Kalliopi I Pappa
author_sort Anastasia Mortaki
collection DOAJ
description <b>Background/Objectives</b>: This study evaluates the diagnostic performance of DNA methylation testing, alone and in combination with cervical cytology, for high-grade squamous intraepithelial lesion (HSIL) detection. <b>Methods</b>: A prospective study was conducted on 170 high-risk HPV (hr-HPV)-positive women. DNA methylation (QIAsure<sup>®</sup>) and cervical cytology were performed prior to cervical large loop excision of the transformation zone (LLETZ). Sensitivity, specificity, and area under the curve (AUC) metrics were calculated, including stratified analyses for HPV16/18 and other hr-HPV genotypes. <b>Results</b>: DNA methylation alone achieved a sensitivity of 69.7%, specificity of 79%, and an AUC of 0.796 for HSIL detection. The combination of cervical cytology and DNA methylation improved sensitivity to 94.7%, specificity to 76.3%, and AUC to 0.860. Stratification by HPV genotype revealed that in HPV16/18-positive cases, DNA methylation alone reached an AUC of 0.790, while the combination with cytology significantly enhanced performance to 0.917. DNA methylation alone demonstrated an AUC of 0.744 for other hr-HPV types, and the combined approach achieved an AUC of 0.849. Specificity for the combined approach was notably higher in HPV16/18-positive women (88.9%) than in other hr-HPV cases (72.4%), whereas the sensitivity of the combined approach was significantly higher in both groups (94.5% vs. 95%, respectively). <b>Conclusions</b>: The integration of DNA methylation with cervical cytology significantly enhances diagnostic accuracy for CIN2+ lesions, especially in HPV16/18-positive cases. However, the comparatively lower AUC and specificity observed in other hrHPV types suggest the need for further optimization to enhance accuracy in non-16/18 infections. These findings support the integration of methylation-based testing with cytology as a valuable triage strategy for improving cervical cancer screening and patient management.
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spelling doaj-art-e3df8751b1174cd5b6f968a7edd685e02025-08-20T02:33:30ZengMDPI AGBiomedicines2227-90592025-05-01135113910.3390/biomedicines13051139The Role of HPV Genotyping, Cytology, and Methylation in the Triage of High-Risk HPV-Positive PatientsAnastasia Mortaki0Athanasios Douligeris1Maria-Anastasia Daskalaki2Eleni-Sivylla Bikouvaraki3Eirini Louizou4George Daskalakis5Alexandros Rodolakis6Themos Grigoriadis7Kalliopi I Pappa81st Department of Obstetrics and Gynecology, National and Kapodistrian University of Athens, “Alexandra” General Hospital, 11528 Athens, Greece1st Department of Obstetrics and Gynecology, National and Kapodistrian University of Athens, “Alexandra” General Hospital, 11528 Athens, Greece1st Department of Obstetrics and Gynecology, National and Kapodistrian University of Athens, “Alexandra” General Hospital, 11528 Athens, GreeceLaboratory of Cell and Gene Therapy, Centre of Basic Research, Biomedical Research Foundation of the Academy of Athens (BRFAA), 11527 Athens, GreeceDepartment of Cytogenetics and Molecular Genetics, Bioiatriki, Group of Health Sciences, 11528 Athens, Greece1st Department of Obstetrics and Gynecology, National and Kapodistrian University of Athens, “Alexandra” General Hospital, 11528 Athens, Greece1st Department of Obstetrics and Gynecology, National and Kapodistrian University of Athens, “Alexandra” General Hospital, 11528 Athens, Greece1st Department of Obstetrics and Gynecology, National and Kapodistrian University of Athens, “Alexandra” General Hospital, 11528 Athens, Greece1st Department of Obstetrics and Gynecology, National and Kapodistrian University of Athens, “Alexandra” General Hospital, 11528 Athens, Greece<b>Background/Objectives</b>: This study evaluates the diagnostic performance of DNA methylation testing, alone and in combination with cervical cytology, for high-grade squamous intraepithelial lesion (HSIL) detection. <b>Methods</b>: A prospective study was conducted on 170 high-risk HPV (hr-HPV)-positive women. DNA methylation (QIAsure<sup>®</sup>) and cervical cytology were performed prior to cervical large loop excision of the transformation zone (LLETZ). Sensitivity, specificity, and area under the curve (AUC) metrics were calculated, including stratified analyses for HPV16/18 and other hr-HPV genotypes. <b>Results</b>: DNA methylation alone achieved a sensitivity of 69.7%, specificity of 79%, and an AUC of 0.796 for HSIL detection. The combination of cervical cytology and DNA methylation improved sensitivity to 94.7%, specificity to 76.3%, and AUC to 0.860. Stratification by HPV genotype revealed that in HPV16/18-positive cases, DNA methylation alone reached an AUC of 0.790, while the combination with cytology significantly enhanced performance to 0.917. DNA methylation alone demonstrated an AUC of 0.744 for other hr-HPV types, and the combined approach achieved an AUC of 0.849. Specificity for the combined approach was notably higher in HPV16/18-positive women (88.9%) than in other hr-HPV cases (72.4%), whereas the sensitivity of the combined approach was significantly higher in both groups (94.5% vs. 95%, respectively). <b>Conclusions</b>: The integration of DNA methylation with cervical cytology significantly enhances diagnostic accuracy for CIN2+ lesions, especially in HPV16/18-positive cases. However, the comparatively lower AUC and specificity observed in other hrHPV types suggest the need for further optimization to enhance accuracy in non-16/18 infections. These findings support the integration of methylation-based testing with cytology as a valuable triage strategy for improving cervical cancer screening and patient management.https://www.mdpi.com/2227-9059/13/5/1139human papillomavirushigh-grade squamous intraepithelial lesionsDNA methylationFAM19A4hsa-miR124-2cervical cytology
spellingShingle Anastasia Mortaki
Athanasios Douligeris
Maria-Anastasia Daskalaki
Eleni-Sivylla Bikouvaraki
Eirini Louizou
George Daskalakis
Alexandros Rodolakis
Themos Grigoriadis
Kalliopi I Pappa
The Role of HPV Genotyping, Cytology, and Methylation in the Triage of High-Risk HPV-Positive Patients
Biomedicines
human papillomavirus
high-grade squamous intraepithelial lesions
DNA methylation
FAM19A4
hsa-miR124-2
cervical cytology
title The Role of HPV Genotyping, Cytology, and Methylation in the Triage of High-Risk HPV-Positive Patients
title_full The Role of HPV Genotyping, Cytology, and Methylation in the Triage of High-Risk HPV-Positive Patients
title_fullStr The Role of HPV Genotyping, Cytology, and Methylation in the Triage of High-Risk HPV-Positive Patients
title_full_unstemmed The Role of HPV Genotyping, Cytology, and Methylation in the Triage of High-Risk HPV-Positive Patients
title_short The Role of HPV Genotyping, Cytology, and Methylation in the Triage of High-Risk HPV-Positive Patients
title_sort role of hpv genotyping cytology and methylation in the triage of high risk hpv positive patients
topic human papillomavirus
high-grade squamous intraepithelial lesions
DNA methylation
FAM19A4
hsa-miR124-2
cervical cytology
url https://www.mdpi.com/2227-9059/13/5/1139
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