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RHD genotyping in weak D phenotypes by multiple polymerase chain reactions
T J Legler1, J H Maas, V Blaschke
1Department of Transfusion Medicine, University of Göttingen, Germany.
Transfusion
|June 20, 1998
Summary
Genetic variations in weak D phenotypes are uncommon. Testing seven RHD gene regions found genetic diversity in only 1.1% of weak D cases, highlighting the rarity of genomic basis in weak D.
Area of Science:
- Genetics
- Immunology
- Molecular Biology
Background:
- Weak D phenotypes represent quantitative variations in D antigen expression.
- The precise genomic underpinnings of weak D phenotypes have remained a subject of scientific debate.
Purpose of the Study:
- To investigate the genomic basis of weak D phenotypes.
- To evaluate various molecular methods for accurate weak D genotyping.
Main Methods:
- Utilized sequence-specific polymerase chain reactions (SSP-PCR) on RHD gene exons 2, 5, and 7.
- Compared SSP-PCR results with intron 4 amplification and serological testing.
- Applied SSP-PCR to the 3' non-coding region for genotyping 94 weak D samples.
Main Results:
- Achieved concordant results between genotyping and phenotyping methods in 201 D-positive and 145 D-negative donors.
- Identified four weak D samples misclassified as D-negative by intron 4 amplification and exon 5 SSP-PCR.
- Discovered a point mutation (T-->G at nucleotide 667) causing a Phe223Val substitution, leading to a false-negative result in one weak D case.
Conclusions:
- Genetic diversity within weak D phenotypes is infrequent.
- Only 1.1% (1 of 90) of true weak D phenotypes exhibited genetic variation across seven tested RHD gene regions.