DNA analysis with BeadChip format, combined with computerized data entry and analysis, permits the prediction of minor blood group antigens.
It has been shown that microarray technology can be used to type DNA and detect new alleles in donor cohorts.
Background : Modern bioarray chip technology enables rapid allele determination in a large number of donors by DNA typing when compared to classical serological methods. We analyzed five single nucleotide polymorphisms (SNPs) associated with Dombrock (DO) alleles in blood samples from donors of different ethnic groups, and found new genetic variants. Method : The BeadChip™ technology used primers for a single multiplex PCR reaction, and allele-specific oligonucleotides with variable 3′-terminal nucleotide for elongation-mediated multiplex analysis (eMAPTM) of the DO polymorphisms. Color-encoded beads displaying the elongation probes were assembled into planar arrays of small footprint on semiconductor chips, permitting the instant imaging of fluorescent elongation products from the entire array. Five SNPs of DO blood group antigens DOA/DOB (nt 378 C>T, 624 T>C, 793 A>G), Hy+/Hy− (nt 323 G>T) and Jo(a+)/Jo(a−)(nt 350 C>T) were analyzed simultaneously. Blood samples from a total of 445 donors, comprising of 100 Thai, 69 Jewish, 58 Chinese and 218 random New York City donors were analyzed. Results : As shown in the table below, the predominant diploid allelic combinations observed in all the groups were DOB/DOB followed by DOA/DOB and then DOA/DOA. In the random donor group, we also observed several less prevalent allele combinations: DOB/HY (14.2%), DOB/JO (1.4%), DOA/HY (0.5%), DOA/JO (2.3%), HY/HY (1.4%), JO/JO(1.8%), and HY/JO (0.9%). Interestingly, two novel alleles in trans with the known alleles: DOB/HA (11% in Thai, 2.9% in Jewish, 6.9% in Chinese and 2.8% in random donors), DOA/HA (1.8% in random donors, 1.7% in Chinese and 1.4% in Jewish), DOB/SH (0.5% in random donors) and HY/SH (1.4 % in random donors) were also detected. HA allele is 378T, 624T, 793A (like JO but with 350C). Hemagglutination studies showed that HA, as predicted from the nucleotide at position 793, encoded Doa antigen. SH allele is 378C, 624C, 793C (like HY but with 323G) and is predicted to encode Dob antigen. Conclusion: The BeadChip™ technology facilitated the identification of novel genetic variants, in addition to accelerating blood group typing. Most prevalent DO allele combinations (in %) Ethnic Group DOB/DOB DOA/DOB DOA/DOA Thai 73 14 2 Jewish 52.2 30.4 9 Chinese 79.3 12.1 0 Random 38.5 21.6 11
Background: Transfusion dependent patients as those with Sickle Cell Disease (SCD) patients become alloimmunized and have the potential to form additional antibodies with such frequency that antigen-negative blood is preferred to prevent further alloimmunization. Blood group genotyping is playing a supporting role in the routine blood banks, especially for provision of antigen-matched blood for these patients. However, current techniques for genomic typing are all labor-intensive and require manual set up and analysis by gel electrophoresis. As a result, DNA microarrays are being developed for the single nucleotide polymorphisms (SNPs) detection in the blood group genes to provide a fast procedure and an automated analysis of numerous blood group polymorphisms. We evaluated the usefulness of DNA microarray to provide a means to precisely match donor blood to the antigen-negative type of SCD patients. Method: A total of 12 DNA samples from patients with SCD (homozygous for HbS) and 84 DNA samples from blood donors, were analyzed by the HEA Beadchip (Hashmi et al, 2005) containing a total of 18 SNPs (FYA/B, FY-GATA, FY265, DOA/B (nt 378, 624, 793), COA/B, LWA/B, DIA/B, SC1/SC2, M/N, S/s, LUA/B, KEL1/2, JKA/B, DO323, DO350, HgbS) in a single reaction. Results: A genotype result was obtained for all SNPs tested on 96 samples within 4 hours of the start of testing. Results obtained by Beadchip analysis in donors were used to provide antigen-matched blood for FYA/B, FY-GATA, FY265, DOA/B, M/N, S/s KEL1/2, JKA/B, for all 12 SCD patients. This technology provided a fast procedure and facilitated the transfusion support with antigen-matched blood in SCD patients allowing the reduction of alloimunization to blood group antigens. Conclusion: This high-throughput DNA analysis has the potential not only to increase the inventory of antigen-negative blood but also to facilitate the matching of RBC component to the recipient’s blood type. It also contributes to the management of transfusions in SCD patients by allowing a more accurate selection of donor units. The application of microarray technology in transfusion medicine may have a tremendous impact on further improvement of the safety of blood transfusion.
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