RHD genotyping in weak D phenotypes by multiple polymerase chain reactions

Transfusion. 1998 May;38(5):434-40. doi: 10.1046/j.1537-2995.1998.38598297211.x.

Abstract

Background: Weak D phenotypes involve a quantitative variation of D. The genomic basis in weak D has been disputed, however.

Study design and methods: Five sequence-specific polymerase chain reactions (SSP-PCRs) on exons 2, 5, and 7 of the RHD gene were evaluated in 248 white and 98 Japanese blood donors and compared with the results obtained by amplification of intron 4 and serology. All methods and SSP-PCR testing on the 3' non-coding region of the RHD gene were applied to the genotyping of 94 DNA samples derived from individuals expressing weak D phenotypes.

Results: Concordant results were obtained with all genotyping and phenotyping methods in testing 201 D-positive and 145 D-negative donors. Four of 94 weak D samples were typed as D-negative by amplification of intron 4 and SSP-PCR on exon 5. Phenotyping with monoclonal antibodies revealed a DVI category in one of these cases and DFR phenotype in three of these cases. One weak D sample, which reacted like normal D-positive cells with all applied monoclonal antibodies, was typed falsely negative by SSP-PCR on exon 5 because of a point mutation at nucleotide 667 (T-->G) that resulted in a Phe223Val amino acid substitution. In this individual, heterozygosity was found at two other amino acid positions (Glu233Gln and Val238Met) by restriction fragment length polymorphism analysis.

Conclusion: Genetic diversity in weak D phenotypes is rare. Only 1 of 90 true weak D phenotypes (1.1%) had a genetic variation in testing on seven gene regions of the RHD gene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Asian Continental Ancestry Group / genetics
  • DNA / analysis
  • European Continental Ancestry Group / genetics
  • Exons
  • Genotype
  • Humans
  • Introns
  • Phenotype
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Single-Stranded Conformational
  • Rh-Hr Blood-Group System / genetics*

Substances

  • Rh-Hr Blood-Group System
  • DNA