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Development of a PCR‐based diagnostic assay for the determination of KEL genotype in donor blood samples
Author(s) -
Murphy M. T.,
Fraser R. H.,
Goddard J. P.
Publication year - 1996
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.1365-3148.1996.d01-62.x
Subject(s) - genotype , microbiology and biotechnology , restriction enzyme , restriction fragment length polymorphism , polymerase chain reaction , biology , phenotype , antigen , restriction site , chemistry , genetics , dna , gene
The polymorphism which determines expression of Kell and Cellano antigens on the red‐cell surface has been reported to be a single C→T nucleotide substitution at residue 701 where T codes for the presence of Kell antigen. This was confirmed by the direct automated sequencing of PCR products amplified from individuals of known Kell phenotype. The substitution creates a Bsm I restriction enzyme site and this has formed the basis for the development of a PCR‐based diagnostic assay for the determination of Kell phenotype in samples of donor blood. The assay is based on RFLP analysis of coamplified PCR products, one of which spans the K/k polymorphic site, and one control fragment which contains a Bsm I site. Digestion of the PCR products with Bsm I restriction enzyme and subsequent gel analysis of the digest allowed unequivocal determination of the K/k status of all of the samples tested.