RHD genotyping in weak D phenotypes by multiple polymerase chain reactions

被引:21
作者
Legler, TJ
Maas, JH
Blaschke, V
Malekan, M
Ohto, H
Lynen, R
Bustami, N
Schwartz, DWM
Mayr, WR
Köhler, M
Panzer, S
机构
[1] Univ Gottingen, Dept Transfus Med, D-37075 Gottingen, Germany
[2] Univ Gottingen, Dept Biochem 1, D-37075 Gottingen, Germany
[3] Univ Vienna, Dept Blood Grp Serol, Clin Blood Grp Serol, Vienna, Austria
[4] Fukushima Med Coll, Blood Transfus Serv, Fukushima, Japan
关键词
D O I
10.1046/j.1537-2995.1998.38598297211.x
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
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' noncoding 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 D-VI 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.
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收藏
页码:434 / 440
页数:7
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