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DNA Reference Reagents for Genotyping RH Variants. | LitMetric

DNA Reference Reagents for Genotyping RH Variants.

J Mol Diagn

Office of Blood Research and Review, Center for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, Maryland. Electronic address:

Published: June 2024

AI Article Synopsis

  • - Patients with Rhesus (RH) blood group variants may need compatible blood transfusions, but existing serologic tests often struggle to identify these variants, necessitating the use of red blood cell genotyping assays for better management of transfusions.
  • - The study developed and validated eight lyophilized DNA reference reagents that represent 21 polymorphisms in the RHD and RHCE genes, utilizing immortalized B-lymphoblastoid cell lines and involving 17 laboratories to assess their accuracy through various molecular assays.
  • - The genotyping results showed a high agreement rate (99.4%) for target RH polymorphisms, validating these DNA reagents as effective reference standards for RH blood group genotyping assay development

Article Abstract

Patients who carry Rhesus (RH) blood group variants may develop Rh alloantibodies requiring matched red blood cell transfusions. Serologic reagents for Rh variants often fail to specifically identify variant Rh antigens and are in limited supply. Therefore, red blood cell genotyping assays are essential for managing transfusions in patients with clinically relevant Rh variants. Well-characterized DNA reference reagents are needed to ensure quality and accuracy of the molecular tests. Eight lyophilized DNA reference reagents, representing 21 polymorphisms in RHD and RHCE, were produced from an existing repository of immortalized B-lymphoblastoid cell lines at the Center for Biologics Evaluation and Research/US Food and Drug Administration. The material was validated through an international collaborative study involving 17 laboratories that evaluated each DNA candidate using molecular assays to characterize RHD and RHCE alleles, including commercial platforms and laboratory-developed testing, such as Sanger sequencing, next-generation sequencing, and third-generation sequencing. The genotyping results showed 99.4% agreement with the expected results for the target RH polymorphisms and 87.9% for RH allele agreement. Most of the discordant RH alleles results were explained by a limited polymorphism coverage in some genotyping methods. Results of stability and accelerated degradation studies support the suitability of these reagents for use as reference standards. The collaborative study results demonstrate the qualification of these eight DNA reagents for use as reference standards for RH blood group genotyping assay development and analytical validation.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11238275PMC
http://dx.doi.org/10.1016/j.jmoldx.2024.02.005DOI Listing

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