J Mol Diagn. 2019 May;21(3):525-537. doi: 10.1016/j.jmoldx.2019.02.003. Epub 2019 Mar 12.
Validated Reference Panel from Renewable Source of Genomic DNA Available for Standardization of Blood Group Genotyping.
Volkova E, Sippert E, Liu M, Mercado T, Denomme GA, Illoh O, Liu Z, Rios M; Collaborative Study Group.
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.
Copyright © 2019 American Society for Investigative Pathology and the Association for Molecular Pathology.
Published by Elsevier Inc. All rights reserved.
PMID: 30872185 DOI: 10.1016/j.jmoldx.2019.02.003