Korean J Clin Pathol.
1999 Apr;19(2):246-251.
Genotyping of Diego Blood Group System by Use of Polymerase Chain Reaction and Nae I Restriction Enzyme
- Affiliations
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- 1Department of Clinical Pathology, Hanyang University College of Medicine, Seoul, Korea.
Abstract
- BACKGROUND
The Diego blood group system consists of two independent pairs of antigens, Dia and Dib. Immunization to Dia or Dib is clinically significant, because anti-Dia and anti-Dib may cause hemolytic transfusion reactions of transfused incompatible red cells or hemolytic disease of the newborn. At the nucleotide level, the difference between the Di a and Di b alleles is a single-base change of exon 19 that results in the substitution of leucine (CTG) for proline (CCG) at position 854.
METHODS
Peripheral blood was collected from 116 patients. DNA was isolated from 50 L of blood. PCR was performed with previously described primers by Bruce et al (S22: 5'-GTC ACGTCGCTCAGCGG, AS13: 5'-GACCTTCCTCCTCATCAA). The 5 L of PCR products were digested by Nae I. We analyzed 10ul of each digested PCR product by electrophoresis on 1.5 % agarose gel with ethidium bromide staining.
RESULTS
A concordance rate of 100 percent was observed between genotyping and phenotyping (105 Di (a-b+), 11 Di (a+b+)).
CONCLUSIONS
This method can be effectively used for the Diego typing and is particularly useful in cases where the serological typing method is difficult as in autoimmune hemolytic anemia.
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