title: Development of a DNA-based genotyping method for the Diego blood group system.
abstract: BACKGROUND
  The paucity of appropriate reagents for serologic typing of the Diego blood group has hindered the identification of the rare Di(b-) blood donors needed to transfuse a Dib antigen-negative patient who presented with anti-Dib. Development of an alternative Di typing approach as a supplement to the current serologic typing method is an important and necessary goal.
  
  
  METHODS
  DI1 and DI2 alleles result from a single C to T substitution at nucleotide 2561 in exon 19 of the human anion exchanger gene causing a proline (DI1) to leucine (DI2) change at amino acid position 854. Allele-specific primers were designed to specifically amplify the DI1 and DI2 alleles using a PCR-based assay system.
  
  
  RESULTS
  A PCR sequence-specific primer (SSP) method for Di genotyping was developed, and the specificity and reproducibility of the method were assessed in a blind control study using serologic tests, family segregation, and DNA sequencing analyses. A total of 1,766 DNA samples from unrelated blood donors were typed for DI1 and DI2 alleles and a single Di(b-) donor was identified. The frequency of DI1 and DI2 alleles among Chinese blood donors was 0.0357 and 0.9643, respectively.
  
  
  CONCLUSIONS
  A simple, accurate, and inexpensive DNA-based PCR-SSP method was established for Di genotyping. The typing results can be visualized on a single photograph within 3 hours, making this reliable method suitable for large-scale typing of potential blood donors without serologic backup.
authors.affiliation: Shen-Zhen Institute of Transfusion Medicine, China. guangwu@szonline.net
authors.forename: Guo-Guang
authors.initials: GG
authors.lastname: Wu
authors.forename: Yu-Qing
authors.initials: YQ
authors.lastname: Su
authors.forename: Qiong
authors.initials: Q
authors.lastname: Yu
authors.forename: Shi-Zheng
authors.initials: SZ
authors.lastname: Jin
authors.forename: Tong-Mao
authors.initials: TM
authors.lastname: Zhao
mesh_terms: D001789:Blood Group Antigens / Q000032:analysis / Q000235:genetics*; D001788:Blood Grouping and Crossmatching / Q000379:methods* / Q000592:standards; D001803:Blood Transfusion / Q000379:methods; D017931:DNA Primers; D005192:Family Health; D005260:Female; D005838:Genotype; D006801:Humans; D008875:Middle Aged; D009101:Multiple Myeloma / Q000628:therapy; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods* / Q000592:standards
chemical_list: D001789:Blood Group Antigens; D017931:DNA Primers