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Dried blood spot-based monitoring of immune and epigenetic biomarkers in occupational exposure studies

Journal of Exposure Science & Environmental Epidemiology (2026)Cite this article

Abstract

Background

Self-sampling of biomaterials is an important means for continuous remote occupational surveillance. Recently, we presented an overview of different self-sampling methods for occupational research. Dried blood spots (DBS), urine, saliva and oral buccal cells were matrices of choice for rapid, minimally invasive sample collection for analysis of a wide range of biomarkers.

Objective

We set up pilot studies to optimise a suitable workflow for minimally invasive matrices in different occupational setups.

Methods

A two-phase pilot study was conducted. In Phase 1, DBS samples were collected from 40 healthy participants according to written and video-recorded protocols. Samples were received intact and without visible damage. We surveyed participants on aspects related to ease of self-sampling and clarity of instructions. In Phase 2, a study of 15 participants, we further refined the protocols to characterise global DNA methylation, oxidative DNA damage (8-OHdG), and immunoarrays (IFN-γ, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, TNF-α, and CRP) in DBS and dried plasma spots (DPS) and assessed the effect of sample storage duration and shipment (stored immediately, and after one, two, four and six weeks).

Results

DBS samples were used to optimise the DNA extraction protocol, which yielded good-quality DNA. We showed the stability for most of the markers up to six weeks. A large number of immune markers could be detected in DBS- and DPS-derived samples, and proteomics was feasible.

Significance

With this study, we confirmed that minimally invasive techniques for good-quality biological sample collection are available for use in future occupational studies.

Impact

This study establishes a robust framework for minimally invasive self-sampling in occupational health research, demonstrating the feasibility of dried blood and plasma spots for high-quality analysis of proteomic, immune, and DNA biomarkers. These methods support scalable, remote, and longitudinal biomolecular surveillance across diverse work environments. Evaluating biomarker stability is essential, as room temperature storage can affect data integrity. Our findings confirm robust proteomic profiling, stable DNA methylation markers, and promising immune marker detection, reinforcing the potential of dried blood spots for reliable long-term health monitoring.

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Fig. 1: Overview of sample collection.
Fig. 2: Transport and storage conditions of collected samples.
Fig. 3: Ease of self-sampling for saliva, urine and dried blood spots (DBS).
Fig. 4: DNA-based markers in finger-prick dried blood spots (fDBS), venous DBS (vDBS), and whole blood (WB).
Fig. 5: Comparison of total protein level among storage conditions.
Fig. 6: Brain derived neurotrophic factor (BDNF) levels (mean ± SD) in venous dried blood spot (vDBS), dried plasma spot (DPS), and plasma (PL) samples.

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Data availability

The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This research was supported by the EPHOR (Horizon 2020 research and innovation programme; grant agreement no. 874703) and INTERCAMBIO (Horizon Europe research and innovation actions; grant agreement no. 101137149) projects. MCT is funded by a Ramón y Cajal fellowship (RYC-2017-01892) from the Spanish Ministry of Science, Innovation and Universities and co-funded by the European Social Fund. ISGlobal acknowledges support from the grant CEX2023-0001290-S funded by MCIN/AEI/ 10.13039/501100011033, and support from the Generalitat de Catalunya through the CERCA Programme. We want to thank the collaborators from Proteomics Leuven - Laboratory of Applied Mass Spectrometry (LAMaS) for their valuable contribution to the research presented in this paper. Parts of this article are based on the following master’s theses: ‘Workflow optimisation of DNA analysis in dried blood spots (DBS)’ by Yanthe Buntinx, KU Leuven, Belgium, and ‘Exploring the Effectiveness of Dried Blood Spot Sampling in Measuring Inflammatory Markers’ by Eva-Maria Hoornaert, KU Leuven, Belgium [57, 58]. The authors would like to thank Laura Smales for their professional language editing and proofreading assistance, which helped improve the clarity and readability of this manuscript.

Funding

This research was supported by the EPHOR (Horizon 2020 research and innovation programme; grant agreement no. 874703) and INTERCAMBIO (Horizon Europe research and innovation actions; grant agreement no. 101137149) projects.

Author information

Author notes

  1. These authors contributed equally: Eva-Maria Hoornaert, Yanthe Buntinx.

  2. These authors jointly supervised this work: Lode Godderis, Manosij Ghosh.

Authors and Affiliations

  1. Department of Public Health and Primary Care, Centre for Environment and Health, Katholieke Universiteit Leuven, Leuven, Belgium

    Evi De Ryck, Eva-Maria Hoornaert, Yanthe Buntinx, Eline Verscheure, Peter HM Hoet, Lode Godderis & Manosij Ghosh

  2. Department of Physiotherapy, Human Physiology and Anatomy, Pain in Motion Research Group (PAIN), Vrije Universiteit Brussel, Brussels, Belgium

    Yanthe Buntinx

  3. Barcelona Institute for Global Health (ISGlobal), Barcelona, Spain

    Michelle C. Turner

  4. Universitat Pompeu Fabra (UPF), Barcelona, Spain

    Michelle C. Turner

  5. CIBER Epidemiología y Salud Pública (CIBERESP), Madrid, Spain

    Michelle C. Turner

  6. Netherlands Organisation for Applied Scientific Research TNO, Risk Analysis for Products in Development, Utrecht, The Netherlands

    Anjoeka Pronk

  7. IDEWE, External Service for Prevention and Protection at work, Heverlee, Belgium

    Lode Godderis

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Contributions

Evi De Ryck: Conceptualisation, methodology, formal analysis, writing—original draft, writing—review and editing, visualisation, supervision, project administration; Eva-Maria Hoornaert: Investigation, writing—review and editing; Yanthe Buntinx: Investigation, writing—review and editing; Eline Verscheure: Investigation, writing—review and editing; Michelle C Turner: Writing—review and editing, funding acquisition; Anjoeka Pronk: Writing—review and editing, funding acquisition; Peter HM Hoet: Supervision, writing—review and editing; Lode Godderis: Supervision, writing—review and editing, funding acquisition; Manosij Ghosh: Conceptualisation, methodology, writing—original draft, writing—review and editing, visualisation, supervision, project administration, funding acquisition.

Corresponding author

Correspondence to Manosij Ghosh.

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The study protocol was approved by the Ethics Committee Research UZ/KU Leuven (S64599).

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De Ryck, E., Hoornaert, EM., Buntinx, Y. et al. Dried blood spot-based monitoring of immune and epigenetic biomarkers in occupational exposure studies. J Expo Sci Environ Epidemiol (2026). https://doi.org/10.1038/s41370-026-00842-1

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