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29 January 2026

Functional DNA Repair Profiling in Translational Medicine: Benchmarking Comet, γH2AX, and NGS Assays Against Clinical Constraints

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1
Department of Pharmaceutical Microbiology and Biochemistry, Medical University of Lodz, 92-215 Lodz, Poland
2
Biobank, Department of Immunology and Allergy, Medical University of Lodz, 92-213 Lodz, Poland
3
Department of Rheumatology, Medical University of Lodz, 92-115 Lodz, Poland
*
Author to whom correspondence should be addressed.
This article belongs to the Special Issue DNA Damage and Repair in Health and Diseases

Abstract

Quantifying DNA repair capacity (DRC) is pivotal for stratifying patients in oncology and autoimmune disorders, yet methodological heterogeneity compromises data reproducibility. While basic research relies on genetically encoded reporters, translational settings demand robust assays compatible with biobanked material, particularly Peripheral Blood Mononuclear Cells (PBMCs). This review benchmarks functional DNA repair assays—ranging from alkaline/neutral comet variants and high-content foci imaging (γH2AX/53BP1) to emerging Next-generation sequencing (NGS)-based break mapping—against the rigors of clinical application. We critically evaluate sensitivity, specificity, and throughput, identifying artifacts introduced by cryopreservation, steroid therapy, and oxidative stress. Furthermore, we propose a “Minimum Reporting Standard” checklist to harmonize DRC quantification. By distinguishing established validation tools from experimental artifacts, this framework aligns assay selection with specific biological endpoints and clinical feasibility.

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