Office of Blood Research and Review, Center for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, Maryland.
Blood Research Institute and Diagnostic Laboratories, Versiti/BloodCenter of Wisconsin, Milwaukee, Wisconsin.
J Mol Diagn. 2019 May;21(3):525-537. doi: 10.1016/j.jmoldx.2019.02.003. Epub 2019 Mar 12.
Extended blood group genotyping is an invaluable tool used for prevention of alloimmunization. Genotyping is particularly suitable when antigens are weak, specific antisera are unavailable, or accurate phenotyping is problematic because of a disease state or recent transfusions. In addition, genotyping facilitates establishment of mass-scale patient-matched donor databases. However, standardization of genotyping technologies has been hindered by the lack of reference panels. A well-characterized renewable reference panel for standardization of blood group genotyping was developed. The panel consists of genomic DNA lyophilized and stored in glass vials. Genomic DNA was extracted in bulk from immortalized lymphoblastoid cell lines, generated by Epstein-Barr virus transformation of peripheral blood lymphocytes harvested from volunteer blood donors. The panel was validated by an international collaborative study involving 28 laboratories that tested each DNA panel member for 41 polymorphisms associated with 17 blood group systems. Overall, analysis of genotyping results showed >98% agreement with the expected outcomes, demonstrating suitability of the material for use as reference. Highest levels of discordance were observed for the genes CR1, CD55, BSG, and RHD. Although limited, observed inconsistencies and procedural limitations reinforce the importance of reference reagents to standardize and harmonize results. Results of stability and accelerated degradation studies support the suitability of this panel for use as reference reagent for blood group genotyping assay development and standardization.
扩展血型基因分型是一种非常有价值的工具，可用于预防同种免疫。当抗原较弱、缺乏特异性抗血清或由于疾病状态或近期输血导致准确表型存在问题时，基因分型特别适用。此外，基因分型有助于建立大规模的患者匹配供体数据库。然而，由于缺乏参考面板，基因分型技术的标准化一直受到阻碍。已经开发出一种经过良好表征的可再生参考面板，用于标准化血型基因分型。该面板由冻干并储存在玻璃小瓶中的基因组 DNA 组成。从从志愿者献血者采集的外周血淋巴细胞中经 EBV 转化生成的永生淋巴母细胞系中批量提取基因组 DNA。该面板通过涉及 28 个实验室的国际合作研究进行了验证，这些实验室对每个 DNA 面板成员进行了 41 种与 17 个血型系统相关的多态性测试。总体而言，基因分型结果的分析表明，与预期结果的一致性>98%，证明该材料适合用作参考。在 CR1、CD55、BSG 和 RHD 等基因中观察到最高水平的不一致。尽管观察到的不一致和程序限制有限，但它们强调了参考试剂对于标准化和协调结果的重要性。稳定性和加速降解研究的结果支持该面板适合用作血型基因分型测定开发和标准化的参考试剂。
