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Review
. 2025 Apr:114:105636.
doi: 10.1016/j.ebiom.2025.105636. Epub 2025 Mar 22.

Reporting of molecular test results from cell-free DNA analyses: expert consensus recommendations from the 2023 European Liquid Biopsy Society ctDNA Workshop

Vincent D de Jager  1 , Patrizio Giacomini  2 , Jennifer A Fairley  3 , Rodrigo A Toledo  4 , Simon J Patton  5 , Simon A Joosse  6 , Claudia Koch  7 , Zandra C Deans  3 ; ELBS ctDNA Workshop Group; Klaus Pantel  7 , Ellen Heitzer  8 , Ed Schuuring  9
Collaborators, Affiliations
Review

Reporting of molecular test results from cell-free DNA analyses: expert consensus recommendations from the 2023 European Liquid Biopsy Society ctDNA Workshop

Vincent D de Jager et al. EBioMedicine. 2025 Apr.

Abstract

The implementation of circulating tumor DNA (ctDNA) in the diagnostic routine may enable non-invasive predictive biomarker testing and treatment optimization in patients who lack a suitable tumor specimen, have failed previous molecular analysis or are clinically ineligible for (re-)biopsy procedures. As the interpretation and reporting are more complex for ctDNA than conventional tissue-based NGS, there is a need for specific guidelines. These will offer support for the reporting of ctDNA test results and will facilitate optimal communication of liquid biopsy findings between diagnostic laboratories and the medical oncology team. Aiming to generate guidelines based on real-world experiences and broad perspectives, we organized a European Liquid Biopsy Society (ELBS) ctDNA workshop, in which forty-four experts and key stakeholders from different molecular diagnostics laboratories, oncology and pathology departments, as well as an IVDR specialist, convened to address significant challenges associated with the reporting of liquid biopsy test results. This report delineates the resulting consensus recommendations for ctDNA test reporting with underlying rationale and background information.

Keywords: Expert consensus recommendations; Liquid biopsy; ctDNA test reporting.

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Conflict of interest statement

Declaration of interests VdJ has received speaker's fees from Roche and Janssen (Johnson & Johnson) (all paid to institution). PG has received payment or honoraria from Illumina for Round Tables on Comprehensive Genomic Profiling, and has a leadership or fiduciary role in an Advisory Board for Thena Biotech. RT has received grants or contracts from AstraZeneca, Beigene pharmaceutics, and Personalis. SP has received financial support from AstraZeneca, MSD, and Johnson & Johnson for EMQN CIC to deliver external quality assessment activities to laboratories worldwide, has received honoraria from AstraZeneca for delivering webinar series, and has received travel costs from AstraZeneca to support delivery of a lecture at a major European conference.SJ has received funding from the EU from the EU4Health call (project name: Building the EU Cancer and Public Health Genomics platform (CAN.HEAL). ES has received unrestricted grants (all paid to UMCG institution) from Abbott, Biocartis, AstraZeneca, Invitae/Archer, Bayer, Bio-Rad, Roche, Agena Bioscience, CC Diagnostics, MSD/MERCK, and SNN/EFRO, has received consulting fees (all paid to UMCG institution) from MSD/Merck, AstraZeneca, Roche, Novartis, Bayer, BMS, Lilly, Amgen, Illumina, Agena Bioscience, CC Diagnostics, Janssen Cilag (Johnson & Johnson), Astellas Pharma, GSK, Sinnovisionlab, Sysmex, and Protyon, has received payments or honoraria (all paid to UMCG institution) from Bio-Rad, Seracare, Roche, Biocartis, Lilly, Agena Bioscience, and Illumina, has received support for attending meetings and/or travel from BioRad, Biocartis, Ageno Sciences, Illumina, Roche/Foundation Medicine, and QCMD, is a board member for the Dutch Society of Pathology (unpaid), European Society of Pathology (unpaid), European Liquid Biopsy Society (unpaid), is a secretary/member of the advisory committee for assessment of molecular diagnostics (cieBOD) (honoraria paid to UMCG institution), is committee member of national guideline advisory (honoraria paid to UMCG institution). CLA has received support for the present manuscript from EU CAN.HEAL (grant n.101080009) (payment to institution). BB has received Oncomine Research Grant from ThermoFisher for analysis of cfDNA in SCLC. AG has received consulting fees from Guardant Health (personal payment) and Foundation Medicine (personal payment). TJNH has received grants or contracts from Roche, BMS, and AstraZeneca (all payments to institution). MH has received consulting fees from Guardant Health, AstraZeneca, Boehringer Ingelheim, Bayer, Roche, Novartis, Merck, Incyte, GSK, and Qiagen, has received payment or honoraria for lectures, presentations, speakers bureaus, manuscript writing or educational events from Janssen, Servier, Bayer, and Seagen, and has received support for attending meetings and/or travel from Janssen, Servier, Nayer, and Seagen. SI has received grants or contracts from Menarini Stemline (contract with own institution (UNIPD)), has received honorarium for one lecture from Sysmex Partec Srl, and has received support for attending meetings and/or travel from Roche Italy SpA (invitation to a Roche sponsored meeting). LK has received payment for expert testimony from AstraZeneca (payment to institution), and has received support for attending meetings and/or travel from AstraZeneca and Roche. MK has received institutional research grants from Roche and Novartis, has received advisory board/consultancy fees from Janssen, Roche, Bayer, Guardant Health, and Zai Lab, has received payment or honoraria for speakers bureaus from Janssen and Roche, and has received support for travel expenses from Janssen, Roche, and Zai Lab. ML has received grants or contracts from KWF Dutch cancer society (payments to institution), has received consulting fees from GlaxoSmithKline B.V. (payments to institution)and Janssen-Cilag B.V. (payments to institution), and AstraZeneca (payments to institution), and has received payments or honoraria for lectures, presentations, speakers bureaus, manuscript writing or educational events from AstraZeneca, Janssen, and Roche (payments to institution) and from AstraZeneca, Janssen, MSD, and Pfizer (payments to institution). MMV has received grants or contracts from AstraZeneca (research contract to own institution), Sumitomo (research contract to own institution), and Merck KGaA (research contract to own institution), and has received payment or honoraria for lectures, presentations, speakers' bureaus, manuscript writing or educational events from The Ricky Rubio Foundation and IX Spanish Symposium of Liquid Biopsy. KN has other financial or non-financial interests as employee of LGC Clinical Diagnostics. SO has received support for the present manuscript from EU project CAN.HEAL (grant n.101080009), German Research Foundation (DFG) DFG_GZ: OS647/18-1, and DFG DFG_GZ: OS647/14-1, has received grants or contracts from DFG DFG_GZ: OS 647/18-1 (research grant), DFG DFG_GZ: OS 647/14-1 (research grant), European Union's H2020: Can.Heal, European Union's H2020: Melcaya, DFG DFG_OS 647/13-1, DFG DFG_OS 647/7-1, Stiftung Deutsche Krebshilfe: HerediVar, Network of University Medicine for Covid-19 (NUM) GenSurv, and DFG DFG_OS 647/1-1 (all research grants), has received payment or honoraria from Illumina Inc. for presentation at GfH Conference, has received support for attending meetings and/or travel from Illumina Inc. for GfH Conference (travel, hotel, conference fees), and Oxford Nanopore Technologies (travel, hotel, conference fees), and has patents planned, issued or pending (PCT/EP2023/061,370, not related to ctDNA testing). AO has received payments or honoraria for lectures, presentations, speakers bureaus, manuscript writing or educational events from Illumina, Inc. and Stemline Therapeutics B.V., has received support for attending meetings and/or travel from Illumina, Inc., and has participated on a Data Safety Monitoring Board or Advisory Board for Illumina, Inc. ER has received grants or contracts from AstraZeneca, Roche Diagnostics, Clovis, GSK, and BMS, has received consulting fees from AstraZeneca, Roche Diagnostics, Clovis, GSK, and BMS, and has received support for attending meetings and/or travel from AZ and BMS. AR has received a Clinical Scientist Fellowship from Cancer Research UK, has received Horizon Europe Guarantee Funding (PANCAID) from United Kingdom Research and Innovation, has a leadership or fiduciary role as and has received honoraria as Director of Biology Research, ACED from Alliance for Cancer Early Detection, and has a leadership or fiduciary role in the National Research Committee of the British Association for Plastic and Reconstructive Surgery. HS has received institutional funding of German Cancer Research Center (DKFZ); competitive third-party funding through the German Centres for Lung Disease (DZL) and the German Cancer Consortium (DKTK) from the German Federal Ministry for Education and Research. All other authors reports no conflicts of interest.

Figures

Fig. 1
Fig. 1
Overview of questionnaire answers. For each statement (in bold), experts provided their level of agreement for ctDNA test reporting. On the right of each statement, the distribution of all expert answers is depicted. Dark green represents ‘agree, essential’, light green represents ‘agree, useful’, red represents ‘disagree’, and grey represents ‘no opinion’.
Fig. 1
Fig. 1
Overview of questionnaire answers. For each statement (in bold), experts provided their level of agreement for ctDNA test reporting. On the right of each statement, the distribution of all expert answers is depicted. Dark green represents ‘agree, essential’, light green represents ‘agree, useful’, red represents ‘disagree’, and grey represents ‘no opinion’.
Fig. 2
Fig. 2
Relationship between the limit of blank (LoB), the limit of detection (LoD), and the limit of quantification (LoQ). The solid brown line represents the range of VAFs from blank sample(s), which are used to determine the LoB, which defines the background noise or the highest VAF expected from a control sample. The dotted green line represents the expected distribution of detections rates from positive samples with various VAFs at the LoD95, which defines the lowest VAF that can be reliably detected with 95% confidence. The LoQ is the lowest VAF of a variant that can be both detected and quantified with acceptable precision and accuracy. This means that not only can the analyte be detected, but the assay can also provide a reliable and reproducible measurement of its concentration. Adapted from Armbruster and Pry (2008). Created with www.BioRender.com.
Fig. 3
Fig. 3
Four examples (A to D) of a clinical case involving a request for plasma-based predictive molecular profiling. Each example includes a typical molecular result, an interpretation of the NGS findings referencing the ELBS recommendations for reporting, and a sample report (Supplementary Files S3, S4, S5 and S6).
See this image and copyright information in PMC

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