A modular verification and validation framework for hybrid-capture next-generation sequencing HLA typing in clinical...

Hybrid-capture next-generation sequencing HLA workflows are increasingly used in clinical histocompatibility laboratories for high-resolution HLA typing. Compared with amplicon-based approaches, they may provide broader coverage of HLA genes, improved ambiguity resolution and the possibility of including additional targets; however, their greater analytical, bioinformatic and interpretative complexity requires a clear definition of intended use, reportable scope and required validation level. A practical and modular framework was developed for the verification and validation of hybrid-capture next-generation sequencing HLA workflows, applicable to IVDR-regulated methods used according to the manufacturer's instructions for use, locally adapted IVDR workflows, research-use-only workflows converted to clinical use, laboratory-developed or laboratory-adapted workflows and outputs not fully covered by manufacturer claims. Core modules required for clinical HLA typing are distinguished from additional modules activated by specific claims or local adaptations, including analytical performance, bioinformatic validation, risk management and reporting governance. It provides an operational pathway to document fitness for intended use and to distinguish clinically reportable results from technically available outputs not validated for diagnostic use. A dataset generated under routine conditions using a commercial full-length HLA capture-based workflow is used as a practical implementation example. A modular structure integrating analytical performance, bioinformatic control, risk-based thinking, reporting governance and continuous monitoring is provided by this framework in clinical immunogenetics and histocompatibility laboratories.