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Updated: Sep 25, 2026
Ultra-Fast Amplicon-Based Next-Generation Sequencing in Non-Squamous Non-Small Cell Lung Cancer
Published on: September 8, 2023
Molecular HPV Detection in Sentinel Lymph Nodes Using Next Generation Sequencing: Concordance with Ultrastaging and
Sara Bønløkke1,2,3, Sara Elizabeth Sponholtz4,5, Pernille Tine Jensen6,7
1Department of Clinical Medicine, Aarhus University, Aarhus N, Denmark. sarasim@clin.au.dk.
Background:
Ultrastaging of sentinel lymph nodes (SLNs) is standard for nodal assessment in early stage cervical cancer but is limited by sampling constraints and workload. Molecular detection of human papillomavirus (HPV) may identify metastatic disease. This study evaluated next generation sequencing (NGS)-based HPV detection in SLNs as a complementary approach to SLN assessment.
Patients And Methods:
This prospective cohort study included 122 women with early stage cervical cancer treated with primary surgery and SLN mapping. SLNs were assessed by ultrastaging and HPV detection using an NGS-based HPV genotyping panel. A subset of patients additionally underwent polymerase chain reaction (PCR)-based HPV analysis.
Results:
A total of 17 patients (13.9%) had SLN metastases identified by ultrastaging, all of which were HPV-positive by NGS. HPV read counts correlated with SLN metastasis size (Spearman's ρ = 0.73, p < 0.001). NGS additionally identified HPV in 13 patients (10.7%) with ultrastaging-negative SLNs. Compared with patients negative by both methods, these patients had significantly larger tumors (p = 0.009) and higher International Federation of Obstetrics and Gynecology (FIGO) stage (p = 0.02), while exhibiting significantly lower HPV read counts than patients positive by both ultrastaging and NGS (p = 0.03). PCR detected HPV in more SLNs than NGS.
Conclusions:
NGS detected HPV in all ultrastaging-confirmed SLN metastases and identified an additional subgroup of patients with molecular HPV detection despite negative ultrastaging. The association between HPV quantity, metastasis size, and adverse clinicopathological characteristics supports the biological relevance of quantitative HPV detection in SLNs. These findings warrant further evaluation of NGS as a complementary molecular biomarker for SLN assessment in larger prospective studies.
