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RHD genotyping in weak D phenotypes by multiple polymerase chain reactions
Author(s) -
Legler T.J.,
Maas J.H.,
Blaschke V.,
Malekan M.,
Ohto H.,
Lynen R.,
Bustami N.,
Schwartz D.W.M.,
Mayr W.R.,
Kohler M.,
Panzer S.
Publication year - 1998
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1046/j.1537-2995.1998.38598297211.x
Subject(s) - genotyping , polymerase chain reaction , phenotype , genetics , biology , genotype , gene
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.