PCR screening for common weak D types shows different distributions in three Central European populations.
BACKGROUND
DNA sequencing showed RHD mutations for all weak D phenotypes investigated in a study from Southwestern Germany. Molecular classification of weak D offers a more reliable basis than serotyping and is relevant for optimal D transfusion strategies.


METHODS
Sequence-specific primers were designed to detect weak D types 1 to 5 and the partial D phenotype HMi in a modular set for conventional PCR analysis. Alternatively, all reactions were multiplexed into a single tube, and the products were identified after automated capillary electrophoresis by their size and fluorescence. Weak D phenotype samples from 436 donors in the Tyrol (Austria) and Northern Germany were investigated by PCR.


RESULTS
More than 90 percent of the weak D types identified by PCR represented type 1, 2, or 3. The distribution among the common types varied between the Tyrol and Northern Germany (p<0.0001). Three new RHD alleles were identified.


CONCLUSIONS
A PCR method of detecting the common weak D types was validated. This PCR system introduces a simple and rapid tool for routine DNA typing of weak D samples. The results confirmed that all weak D phenotype samples identified by current serologic criteria carry altered D proteins.
Müller
T H
TH
Oldenburg Institute, German Red Cross Blood Donor Service NSOB, Oldenburg, Germany. tmueller@rdg.behringer-ingelheim.com
Wagner
F F
FF
Trockenbacher
A
A
Eicher
N I
NI
Flegel
W A
WA
Schönitzer
D
D
Schunter
F
F
Gassner
C
C
D000483:Alleles; D001317:Austria; D017931:DNA Primers; D005858:Germany; D006801:Humans; D007519:Isoantigens / Q000235:genetics*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012204:Rh-Hr Blood-Group System / Q000235:genetics / Q000276:immunology*
D017931:DNA Primers; D007519:Isoantigens; D012204:Rh-Hr Blood-Group System