Genotyping of Diego Blood Group System by Use of Polymerase Chain Reaction and Nae I Restriction Enzyme / 대한임상병리학회지

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.