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Genetic changes in D weak Spanish donors
Author(s) -
Torres P.,
García F.,
Jurado M.,
Calle C.,
Rodriguez M. A.
Publication year - 2000
Publication title -
transfusion medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.471
H-Index - 59
eISSN - 1365-3148
pISSN - 0958-7578
DOI - 10.1046/j.1365-3148.2000.00261-29.x
Subject(s) - exon , microbiology and biotechnology , gene , genetics , biology , mutation , nucleotide , gene mutation
I ntroduction Recently some mutations were described in RHD gene associated to D weak. We have studied these changes in our blood donors. M aterial and M ethod 56 D weak samples from our donors were studied in the following way: DNA extraction with DNAzol, PCR‐SSP of some fragments of RHD gene and digestion with Alw44I, Sac I, Alu l, Msp l restriction enzymes (F. Wagner, Blood 1999; 93: 385–393). In two samples with fenotipe ce, exons 4 and 5 of gene RHD were sequenced. In 6 samples, which seemed to be antigen D variant, all the exons of the RHD gene were amplified by PCR‐SSP. We also sequenced exons 4, 5 and 7 of the RHD gene and exon 5 of RHCE gene in one case. In two other cases, DVl atipical variants, RNA was extracted from reticulocytes and then cDNA of RHD gene was sequenced. R esults 28 samples have fenotype cEe and in all of them the mutation described in nucleotide 1154(G‐C) was detected. Of 17 samples with fenotype Cce 15 showed the described mutation in nucleotide 809(T‐G) and the remaining 2 showed a mutation in nucleotide 8(C‐G) as was previously pointed out. Two samples have ce fenotype and mutations in nucleotides 602(C‐G) and 667(T‐G). No mutations were located in 3 donors carrying Cce fenotype. Six of our cases belong to antigen D variants; one case is a caucasian cDe donor with a DAR variant with ceAR. Another case, cDEe fenotype, is a type l DVI variant. Two other CcDe fenotypes are DVl atypical variants due to the fact that they present a sustitution of RHD gene exons 3,4 and 5 for those corresponding to the RHCE gene. The study of the last two variants is being completed at present. C onclusions 1. We confirm in our study that D weak is associated to changes, previously described by Wagner et al . in RHD gene. 2. There is a strong correlation between haplotypes and genetic mutations. 3. The most frecuent haplotype associated to D weak in our donors is cDE(R2) showing a mutation in nucleotide 1154. 4. A DAR variant with ceAR has been detected in a caucasian donor. 5. We find ourselves in the position to confirm the existence of a new type of D Vl variant.