Digital droplet PCR-based quantification of ccfHPV-DNA as liquid biopsy in HPV-driven cervical and vulvar cancer

Q: What is the purpose of measuring ccfHPV-DNA in this study?

To evaluate ccfHPV-DNA assessed by ddPCR as a biomarker for molecular therapy monitoring across different stages and disease settings in HPV-driven cervical and vulvar cancer.

Q: How were blood samples processed and how was ccfHPV-DNA measured?

Blood samples were collected before and after therapy, and circulating cell-free HPV DNA was quantified using droplet digital PCR (ddPCR) and correlated with clinical data.

Q: What main results link ddPCR ccfHPV-DNA with treatment response and disease stage?

Mean ccfHPV-DNA copy numbers were higher in pretreatment than post-treatment samples, increased with higher FIGO stages, and were higher in T-stage 2/3 compared with T-stage 1.

Sep 24, 2026

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The study evaluates circulating cell-free HPV DNA (ccfHPV-DNA) quantified by droplet digital PCR (ddPCR) as a biomarker for molecular therapy monitoring in early, advanced, relapsed, and metastatic HPV-driven cervical and vulvar cancer. The work targets HPV16/18-driven disease and relates pre- and post-therapeutic blood ddPCR results with clinical data. Pretreatment samples showed higher copy numbers than post-treatment, with levels increasing with FIGO stage and T-stage 2/3. ddPCR-based ccfHPV-DNA measurement using small plasma volumes appears feasible and may support assessment in novel HPV-directed therapies.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages8

Reading Time20 min

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Abstract

Purpose

Methods

Results

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Introduction

Journal of Cancer Research and Clinical Oncology (2023) 149:12597–12604 [https://doi.org/10.1007/s00432-023-05077-3](https://doi.org/10.1007/s00432-023-05077-3) Digital droplet PCR‑based quantification of ccfHPV‑DNA as liquid biopsy in HPV‑driven cervical and vulvar cancer Fabinshy Thangarajah1,2 · Jana Busshoff2 · Janina Salamon2 · Marie‑Sandrine Pruss2 · Caroline Lenz2 · Bernd Morgenstern2 · Martin Hellmich3 · Hans Anton Schlößer4 · Maximilian Lenz5 · Christian Domröse2 · Michael R. Mallmann2 · Peter Mallmann2 · Jonathan Weiß6 · Fabian Franzen6 · Sabine Merkelbach‑Bruse7 · Elke Binot7 · Marie‑Lisa Eich7 · Reinhardt Büttner7 · Anne Maria Schultheis7 · Christina Alidousty7 Received: 23 May 2023 / Accepted: 29 June 2023 / Published online: 14 July 2023 © The Author(s) 2023 Abstract Purpose More than 99% of cervical cancers and up to 40% of vulvar cancers are human papillomavirus (HPV) related. HPV 16 and 18 are the most relevant subtypes. Novel technologies allow the detection of minimal amounts of circulating cell-free HPV DNA (ccfHPV-DNA) . The aim of this study was to evaluate ccfHPV-DNA assessed by droplet digital PCR (ddPCR) as a biomarker for molecular therapy monitoring in early, advanced, relapsed and metastatic HPV-driven cervical and vulvar cancer. Methods Inclusion criteria of the study were histologically proven HPV 16/18-driven cervical and vulvar cancer with first diagnosed disease, newly diagnosed recurrence, or progression of disease. Blood samples were taken pre-and post-therapeutically. Circulating cell-free HPV DNA was quantified using ddPCR and the results were correlated with clinical data. Results The mean copy number of ccfHPV-DNA was 838.6 (± 3089.1) in pretreatment and 2.3 (± 6.4) in post-treatment samples (p < 0.05). The copy number of ccfHPV-DNA increased with higher FIGO stages (p < 0.05), which are commonly used for clinical staging/assessment. Furthermore, we compared the distribution of copy numbers between T-stage 1 versus T-stage 2/3 . We could show higher copy number level of ccfHPV-DNA in T-stage 2/3 (p < 0.05) . Conclusions Therapy monitoring with determination of ccfHPV-DNA by ddPCR with a small amount of plasma reflects response to therapy and appears feasible for patients in advanced cancer stages of cervical and vulvar cancer. This promising tool should be examined as marker of therapy monitoring in particular in novel HPV-directed therapies. Keywords Liquid biopsy · ddPCR · Cervical cancer · Vulvar cancer · cfDNA · cHPV Anne Maria Schultheis, Christina Alidousty equally contributed. * Fabinshy Thangarajah [Fabinshy.Thangarajah@uk-essen.de](Fabinshy.Thangarajah@uk-essen.de) * Anne Maria Schultheis [Anne.Schultheis@uk-koeln.de](Anne.Schultheis@uk-koeln.de) 1 Department of Gynecology and Obstetrics, University Hospital of Essen, University of Duisburg Essen, Faculty of Medicine, Essen, Germany 2 Department of Obstetrics and Gynecology, University of Cologne, Faculty of Medicine and University Hospital Cologne, Cologne, Germany 3 Institute of Medical Statistics, Informatics and Epidemiology, University of Cologne, Cologne, Germany 4 Center for Molecular Medicine Cologne and Department of General, Visceral and Cancer Surgery, University Hospital of Cologne, Medical Faculty, Cologne, Germany 5 Department for Orthopaedic and Trauma Surgery, University Hospital of Cologne, Medical Faculty, Cologne, Germany 6 Department I of Internal Medicine, Center for Integrated Oncology Aachen Bonn Cologne Duesseldorf, University Hospital of Cologne, Medical Faculty, Cologne, Germany 7 Institute of Pathology, University Hospital of Cologne, Medical Faculty, Cologne, Germany Introduction More than 99% of cervical cancers and up to 40% of vulvar cancers are human papillomavirus (HPV) related (Zhang et al. 2018 ; Schnürch et al. 2015) . The most frequent HPV subtypes are HPV 16 and 18 (Burd 2003) . During the past decades, additional treatment options have improved the

FAQs

What is the purpose of measuring ccfHPV-DNA in this study?

To evaluate ccfHPV-DNA assessed by ddPCR as a biomarker for molecular therapy monitoring across different stages and disease settings in HPV-driven cervical and vulvar cancer.

How were blood samples processed and how was ccfHPV-DNA measured?

Blood samples were collected before and after therapy, and circulating cell-free HPV DNA was quantified using droplet digital PCR (ddPCR) and correlated with clinical data.

What main results link ddPCR ccfHPV-DNA with treatment response and disease stage?

Mean ccfHPV-DNA copy numbers were higher in pretreatment than post-treatment samples, increased with higher FIGO stages, and were higher in T-stage 2/3 compared with T-stage 1.

Sep 24, 2026

Uploaded by Dipper