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- W4387572088 abstract "New alleles are often discovered when targeted genotyping predicts antigen positive and serology shows negative, weak, or variable reactivity. Donor discrepancy resolution is important for correct antigen labeling. We investigated five donor samples with RhCE discrepancy between serology and PreciseType HEA. Serologic testing was done by standard tube methods using Immucor Gamma-clone, Ortho BioClone, Quotient ALBAclone and Bio-Rad Seraclone antisera. DNA was isolated from white blood cells. RHCE BeadChip (Immucor) and Sanger sequencing of RHCE exons 1-10 and flanking intron regions were performed. RHCE*E-specific sequencing (S1, S3) or long-range PCR of RHCE*C followed by allele-specific sequencing (S5) were done to phase variant sequences. Table 1 summarizes the five donors and findings. S1–S4 were predicted C–E+c+e+ by HEA with discrepancies in serologic testing of E (S1–S3) or e (S4) antigens. S1 had variable reactivity with Anti-E reagents (Ortho nonreactive, Immucor weakly reactive, Quotient strongly reactive). RHCE BeadChip confirmed HEA results, identified c.667G >T change (p.Val223Phe) in exon 5, and placed it on ce (ceMO). However, E-specific sequencing found c.667T on cE. S2 showed variable reactivity with Anti-E (Ortho very weakly reactive, Bio-Rad and Immucor nonreactive). RHCE BeadChip found no variant and sequencing identified c.885G >A change (p.Met295Ile) in exon 6. S3 RBCs typed E– with Bio-Rad Anti-E. RHCE BeadChip found no variant, while sequencing revealed c.341G >A change (p.Arg114Gln) in exon 3. E-specific sequencing found c.341A change on cE. S4 RBCs typed e– with Immucor and Ortho Anti-e. No variant was detected by RHCE BeadChip. Sequencing detected c.1115T >A change (p.Leu372Stop) in exon 8. S5 was predicted C+E–c+e+ by HEA, but RBCs typed C− with Ortho Anti-C and reacted weakly with Immucor Anti-C. RHCE BeadChip detected no variant. Sanger sequencing found c.19C >T change (p.Arg7Trp) in exon 1 and c.512A >C change (His171Pro) in Exon 4. Further testing showed c.19T on ce and c.512C on Ce. We report six novel RHCE alleles in five donor samples: cE(667T), cE(885A), cE(341A), ce(1115A), Ce(512C), and ce(19T). The cE(667T) with variable E reactivity is similar to that reported for e on ce(667T), and Ce(667T). The E−/E+vw phenotype associated with cE(885A) is consistent with the weak partial D and Del phenotypes associated with RHD*885A. The c.341A has been reported on ce with weak e and expression of low prevalence JAL antigen. Unfortunately, we were unable to test S3 for JAL. The RHCE*ce(1115A) is presumed null given the premature stop codon and e− typing. Similar to weak c encoded by ce(512G), Ce(512C) encodes weak C. While c.19T on RHD results in weak D, it doesn’t appear to affect c expression on ce, evidenced by strong c+ typing." @default.
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- W4387572088 date "2023-10-01" @default.
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- W4387572088 title "OA3‐AM23‐TU‐12 | Six Novel RHCE Alleles Identified in Investigation of Donor Antigen Typing Discrepancies" @default.
- W4387572088 doi "https://doi.org/10.1111/trf.50_17554" @default.
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