<i>RHCE</i> alleles detected after weak and/or discrepant results in automated Rh blood grouping of blood donors in Northern Germany

Döscher, Andrea; Vogt, Claudia; Bittner, Rita; Gerdes, Ingrid; Petershofen, Eduard K.; Wagner, Franz F.

doi:10.1111/j.1537-2995.2009.02221.x

Cited by 19 publications

(26 citation statements)

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“…In a parallel study by Doescher and colleagues 24 describing RhCE variants in Northern Germany, only 10 of these 22 RHCE alleles were observed, although the variety of alleles was similar. However, none of our donors and patients overlapped with those found in the Northern German study, published in this issue of TRANSFUSION.…”

Section: Resultsmentioning

confidence: 91%

RhCE protein variants in Southwestern Germany detected by serologic routine testing

et al. 2009

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BACKGROUND Variant RHCE alleles with diminished expression of C, c, E, and e antigens have been described and indicate the genetic diversity of this gene locus in several populations. In this study the molecular background of variant RhCE antigens identified by standard serologic routine testing in German blood donors and patients was determined. STUDY DESIGN AND METHODS Samples from blood donors and patients were routinely analyzed for RhCE phenotype using the PK7200 analyzer with two sets of monoclonal anti-C, -c, -E, and -e reagents. Samples with confirmed variant RhCE antigens were analyzed by nucleotide sequencing of the 10 RHCE exons. A multiplex polymerase chain reaction with sequence-specific priming (PCR-SSP) method was established for rapid typing of the rare RHCE alleles. RESULTS We identified 43 samples with serologic RhCE variants. Molecular analysis revealed variant RHCE alleles in 34 samples. Altogether 22 RHCE alleles were detected; 10 have not been published before. Twenty alleles harbored distinct single-nucleotide substitutions, 18 of which encoded amino acid changes and 2 of which occurred in noncoding regions. Two samples represented RHCE-D-CE hybrid alleles involving different segments of the RHCE Exon 5. A multiplex PCR-SSP screening for 17 RHCE alleles was negative in 1344 samples of the DNA bank GerBS. The cumulative phenotype frequency was estimated between 1 in 488 (0.20%) and 1 in 8449 (0.012%). CONCLUSION Single-amino-acid substitutions were the molecular basis for variant RhCE antigen expression in most samples. Nucleotide substitutions in RHCE exons were excluded as possible mechanism of diminished RhCE antigen expression in one-fifth of the serologically identified samples.

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Section: Resultsmentioning

confidence: 91%

RhCE protein variants in Southwestern Germany detected by serologic routine testing

et al. 2009

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“…While this manuscript was being reviewed, Dö scher et al [16] reported that a sample with a weak expression of c had the 662C>G (Pro221Arg) change that we report here. The association with Be a was not realized.…”

Section: Discussionmentioning

confidence: 88%

The low prevalence Rh antigen Be^a(Rh36) is associated withRHCE*ce662C>G in exon 5, which is predicted to encode Rhce 221Arg

et al. 2010

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Background and Objectives The low prevalence antigen, Be a , is produced by a complex that also produces weak c, e and f (ce). We report here the molecular basis associated with Be a antigen expression. Materials and MethodsPeripheral blood samples from four Be(a+) probands were tested. Haemagglutination, gDNA extraction, PCR-based assays, reticulocyte RNA isolation, Rh-cDNA analyses, and sequencing were performed by standard procedures.Results RBCs from Probands 1 and 3 were D)C)E)c+e+, and from Probands 2 and 4 were D+C+E)c+ W e+. In proband 1, cDNA sequencing of RHCE revealed heterozygosity of nucleotide (nt) 662C ⁄ G in exon 5 of RHCE*ce. No other nucleotide changes were observed. As the 662C>G nucleotide change ablates a MscI restriction enzyme cleavage site, PCR-RFLP analysis was performed and the RHCE*ce nt 662C ⁄ G heterozygosity was detected on gDNA from the four probands and two children from both Proband 3 and Proband 4. ConclusionThe low prevalence Rh antigen, Be a , is associated with a single nucleotide change in exon 5 of RHCE*ce; that of 662C>G and this change is predicted to alter proline at amino acid position 221 of Rhce to arginine. The fundamental differences in the properties of these two amino acids may impose a steric and ⁄ or charge-related effect on the protein, and thereby provide an explanation for the weakened expression of c, e and f (ce) antigens in the Be a phenotype.

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“…However, the relevant variation may be located in the promoter region as exemplified by the 'African' Fy(a-b-) phenotype caused by Regrettably, a few inactivating mutations have not been characterized yet: there are reports of absent D [23] or considerably diminished C [24,25] antigens encoded by seemingly normal alleles, precluding a safe 'molecular only' prediction of the respective phenotypes. Another possible pitfall is the presence of extra mutations that are not tested but cause a distinct phenotype [26].…”

Section: Pitfalls In Molecular Antigen Predictionmentioning

confidence: 95%

“…Its great advantage is the detection of any change to the coding sequence. Therefore, sequencing has become a central part of the classification of variant samples that cannot be classified by other methods [24,25].…”

Section: Sequencingmentioning

confidence: 99%

Molecular testing in transfusion medicine

Wagner

2010

Expert Opinion on Medical Diagnostics

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RHCE alleles detected after weak and/or discrepant results in automated Rh blood grouping of blood donors in Northern Germany

Abstract: In addition to RHcE(M167K), a large number of different alleles are underlying CcEe typing problems. Molecular mechanisms parallel those found in RHD. Elucidation of the molecular bases of variant antigens is important to improve serologic and molecular typing methods.

Cited by 19 publications

References 31 publications

RhCE protein variants in Southwestern Germany detected by serologic routine testing

RhCE protein variants in Southwestern Germany detected by serologic routine testing

The low prevalence Rh antigen Be^a(Rh36) is associated withRHCE*ce662C>G in exon 5, which is predicted to encode Rhce 221Arg

Molecular testing in transfusion medicine

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RHCE alleles detected after weak and/or discrepant results in automated Rh blood grouping of blood donors in Northern Germany

Abstract: In addition to RHcE(M167K), a large number of different alleles are underlying CcEe typing problems. Molecular mechanisms parallel those found in RHD. Elucidation of the molecular bases of variant antigens is important to improve serologic and molecular typing methods.

Cited by 19 publications

References 31 publications

RhCE protein variants in Southwestern Germany detected by serologic routine testing

RhCE protein variants in Southwestern Germany detected by serologic routine testing

The low prevalence Rh antigen Bea(Rh36) is associated withRHCE*ce662C>G in exon 5, which is predicted to encode Rhce 221Arg

Molecular testing in transfusion medicine

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The low prevalence Rh antigen Be^a(Rh36) is associated withRHCE*ce662C>G in exon 5, which is predicted to encode Rhce 221Arg