NGS-based detection of circulating HPV DNA offers promise for monitoring for early recurrence of cervical cancer

A study published in Cancer last month showed promising results for an ultrasensitive next-generation sequencing (NGS) approach to detect circulating cell-free human papillomavirus (ccfHPV DNA), demonstrating its prognostic and surveillance value in patients with cervical cancer. The researchers found that levels of ccfHPV DNA were a strong prognostic biomarker for residual disease or recurrence of cervical cancer.

Cervical cancer remains a significant public health challenge, affecting more than 600,000 women globally each year and claiming more than 340,000 lives, according to the World Health Organization. In the U.S., payers and providers grapple with recurrence rates that hover around 12% for early-stage disease and climb to 60% in advanced cases, often leading to dismal five-year survival rates after recurrence of between 25% and 39%.

ccfDNA is fragmented DNA released into the blood by cell death and other processes. Detection of cell-free DNA from tumors is known as circulating tumor DNA. HPV is a cause of cervical cancer, and researchers have started to study whether cell-free DNA of the HPV virus circulating in the blood could be a useful biomarker for tracking cervical cancer.

Traditional monitoring relies on imaging and clinical exams, which can miss minimal residual disease and delay interventions, escalating treatment costs and straining population health resources. For value-based care, clinical adoption of NGS for ccfHPV DNA could emerge as a potential solution, enabling earlier detection and personalized follow-up strategies.

Led by Sara Bønløkke, M.D., Ph.D., from the Department of Clinical Medicine at Aarhus University in Denmark, the researchers conducted a prospective cohort study that enrolled 141 women with International Federation of Gynecology and Obstetrics (FIGO) 2018 stage IA1-IVB cervical cancer between 2018 and 2020. The study aimed to evaluate the prognostic value of ccfHPV DNA in pretreatment samples and to elucidate its role in detecting residual disease and predicting recurrence post treatment.

Participants were stratified by treatment: primary surgery (n = 50), primary surgery plus adjuvant oncology (n = 22), and primary oncological treatment (n = 69, typically chemoradiotherapy). Blood samples were collected before treatment, at the first posttreatment visit (approximately three to four months later) and during a three-year follow-up, with observation extending up to 6.5 years.

The methodology leveraged a custom NGS panel targeting 27 high-risk HPV genotypes classified as carcinogenic by the International Agency for Research on Cancer. Statistical analyses, including Cox proportional hazards models (adjusted for factors such as FIGO stage and tumor size), evaluated associations with recurrence-free survival.

Key findings supported the biomarker's robustness. Pretreatment ccfHPV DNA positivity varied by group — 8% in primary surgery, 36% in surgery plus adjuvant and 83% in primary oncology — and was linked to a nearly fivefold increased recurrence risk. At the first posttreatment sample, positivity emerged as a strong predictor of recurrence (adjusted HR 19.1), with predictive metrics demonstrating a sensitivity of 45%, a specificity of 98%, a positive predictive value (PPV) of 83%, and a negative predictive value (NPV) of 90% for recurrence within 50 months. Longitudinal monitoring revealed that in 61% of the 23 evaluable recurrence cases, ccfHPV DNA became detectable before clinical or imaging evidence, with an average lead time of 144 days and a median of 58 days. Overall posttreatment positivity showed a sensitivity of 61%, a specificity of 96%, a PPV of 78% and an NPV of 92%.

Challenges remain, however, including moderate sensitivity (possibly due to HPV integration in some tumors) and the need for larger, multicenter validation to confirm generalizability. The study faced limitations like sample collection variability amid COVID-19 disruptions, but its funding from nonpharmaceutical sources ensures unbiased insights.

Early detection via ccfHPV DNA could shift paradigms from reactive to proactive care, reducing the need for frequent imaging scans, which can cost between $1,500 and $3,000 per session and expose patients to radiation. Bønløkke and her colleagues add that “larger multicenter studies are needed to validate its clinical utility … into standard care.”