Development of a DNA-based genotyping method for the Diego blood group system.
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.
Wu
Guo-Guang
GG
Shen-Zhen Institute of Transfusion Medicine, China. guangwu@szonline.net
Su
Yu-Qing
YQ
Yu
Qiong
Q
Jin
Shi-Zheng
SZ
Zhao
Tong-Mao
TM
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
D001789:Blood Group Antigens; D017931:DNA Primers