<i>RHCE*01 (</i><scp>c.499A&gt;G</scp>,<scp>p.Met167Val</scp><i>)</i>allele: Weak RhE expression which does not require the E‐specific proline 226

Durieux-Roussel, Elisabeth; Laget, Laurine; Filosa, Lugdivine; Izard, C; Bailly, Pascal; Chiaroni, Jacques

doi:10.1111/trf.16247

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“…In the context of routine immunohematology, serological techniques enable us to characterize most of the Rh phenotypes, and within the last decade, molecular biology solutions have helped compensate for the main limitations of serological techniques [ 12 , 13 , 14 , 15 , 16 , 17 , 18 ]. Still, in some situations, ambiguities or discrepancies cannot be resolved, especially for RhCE, and uncertainties remain regarding the transfusion instructions; in this context, the French Blood Center, whose main mission is to ensure France’s self-sufficiency in blood products under optimal safety and quality conditions, is continuously setting up molecular typing strategies in line with technological developments [ 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 ].…”

Section: Introductionmentioning

confidence: 99%

Resolution of RHCE Haplotype Ambiguities in Transfusion Settings

Izard,

Laget,

Beley

et al. 2024

IJMS

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Red blood cell (RBC) transfusion, limited by patient alloimmunization, demands accurate blood group typing. The Rh system requires specific attention due to the limitations of serological phenotyping methods. Although these have been compensated for by molecular biology solutions, some RhCE ambiguities remain unresolved. The RHCE mRNA length is compatible with full-length analysis and haplotype discrimination, but the RHCE mRNA analyses reported so far are based on reticulocyte isolation and molecular biology protocols that are fastidious to implement in a routine context. We aim to present the most efficient reticulocyte isolation method, combined with an RT-PCR sequencing protocol that embraces the phasing of all haplotype configurations and identification of any allele. Two protocols were tested for reticulocyte isolation based either on their size/density properties or on their specific antigenicity. We show that the reticulocyte sorting method by antigen specificity from EDTA blood samples collected up to 48 h before processing is the most efficient and that the combination of an RHCE-specific RT-PCR followed by RHCE allele-specific sequencing enables analysis of cDNA RHCE haplotypes. All samples analyzed show full concordance between RHCE phenotype and haplotype sequencing. Two samples from the immunohematology laboratory with ambiguous results were successfully analyzed and resolved, one of them displaying a novel RHCE allele (RHCE*03 c.340C>T).

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