Development of a quality control panel based on patient-derived immortalized B-Lymphoblastoid cell lines for thalassemia genotyping.

Background: Thalassemia diagnosis relies on precise genetic testing, which requires reliable quality control (QC) materials. Current options, such as limited clinical samples or synthetic DNA, lack both renewable supply and the ability to authentically mimic patient samples across the entire testing process. To address this gap, we developed and validated a novel panel of QC materials based on patient-derived Immortalized B-Lymphoblastoid cell lines for thalassemia genotyping. Methods: PBMCs from 12 patients with thalassemia were immortalized using an optimized EBV-transformation protocol supplemented with specific growth factors and regulators. The resulting cell lines were then prepared as QC materials by cryopreservation at a defined optimal density in a serum-free solution. We assessed the stability of these QC materials under various conditions, including freeze-thaw cycles, long-term and short-term storage, by analyzing DNA integrity via agarose gel electrophoresis, quantifying Results: The optimized EBV-transformation protocol significantly outperformed the basic method, increasing the transformation efficiency to 90.0% (from 53.1%, Conclusion: We have established a comprehensive and stable QC panel for thalassemia genotyping that serves as a standardized resource for quality assurance, with the potential to be extended as a model for other genetic disorders.