Genotyping of Diego Blood Group System by Use of Polymerase Chain Reaction and Nae I Restriction Enzyme.
- Author:
Duck An KIM
1
;
Think You KIM
;
Tae Yeal CHOI
Author Information
1. Department of Clinical Pathology, Hanyang University College of Medicine, Seoul, Korea.
- Publication Type:Original Article
- Keywords:
Diego blood group;
Genotyping;
PCR-RFLP
- MeSH:
Alleles;
Anemia, Hemolytic, Autoimmune;
Blood Group Incompatibility;
DNA;
Electrophoresis;
Ethidium;
Exons;
Humans;
Immunization;
Infant, Newborn;
Leucine;
Polymerase Chain Reaction*;
Proline;
Sepharose
- From:Korean Journal of Clinical Pathology
1999;19(2):246-251
- CountryRepublic of Korea
- Language:Korean
-
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.
