ObjectivesThis study aimed at identifying new ABO alleles from155 unrelated blood samples with potential ABO discrepancy in a Chinese Han population of 835 144 donors.BackgroundSerological strategies and genotyping are crucial for the precise determination of ABO discrepancy.MethodsTheir ABO phenotypes and plasma glycosyltransferase activity were determined by standard forward and reverse typing and dilution tests. The genomic DNA of the ABO gene was amplified by polymerase chain reaction and sequenced. The frequency of ABO subgroup alleles associated with ABO discrepancy was analysed.ResultsSerological analysis indicated that 53, 96 and 6 samples with ABO discrepancy were identified in the A, B and O subgroups, respectively. Genetic analysis revealed 12 novel alleles among the 46 associated with serologic ABO discrepancy. The majority of novel alleles was obtained from point mutations or single base insertion in Exons 6 to 7 of the ABO gene. The most frequent alleles were ABO*cisAB.01 (14/53, 26.42%) and ABO*A2.05 (7/53, 13.2%) in the A subgroup and ABO*BA.02 (34/96, 35.42%) and ABO*BEL.11 (15/96, 15.62%) in the B subgroup. Samples with the same ABO subgroup allele displayed different phenotypes, such as ABO*AX.13, ABO*BW.03, ABO*BW.12, ABO*BW.15, ABO*BEL.03, ABO*BEL.10 and ABO*BEL.11.ConclusionThis study identified 12 novel alleles among the 46 associated with serologic ABO discrepancies. ABO genotyping is needed for the accurate evaluation of blood phenotype to improve the safety of blood transfusion.
Background: Some blood groups, such as S and s blood groups in the MNS blood group system, and Kidd and CTL2 blood group systems, can cause severe fetal and newborn alloimmune disorders. Non-invasive prenatal testing (NIPT) to predict fetal blood groups and knowledge of local blood group gene frequency are both important for pregnancy management decisions. Droplet digital PCR (ddPCR) has high specificity and sensitivity in detecting fetal single nucleotide variation. Objectives: The objective is to predict fetal Ss, Kidd, and CTL2 blood groups using multiplex ddPCR. The gene frequencies of three blood groups were detected by ddPCR in northwest China. Design: This is a prospective study. Methods: Cell-free fetal DNA isolated from 26 healthy single pregnant women at different gestational stages was tested with QX200 Droplet Digital PCR. Results were compared with fetal genotypes. DNA samples purified from 20 blood pools containing a total of 1000 donors in northwest China were subjected to ddPCR to detect the gene frequency of three blood groups. Results: Ss, Kidd, and CTL2 blood groups of 26 pregnant fetuses were accurately detected by multiplex ddPCR. The multiplex ddPCR results were consistent with the Sanger sequencing results of 26 fetal blood samples after birth. The gene frequencies of the three blood groups detected by ddPCR were 9.30% for S, 90.70% for s, 48.43% for Jka, 51.57% for Jkb, 66.57% for HNA-3A, and 33.43% for HNA-3B. Conclusions: It is reliable to predict fetal Ss, Kidd, and CTL2 blood groups by multiplex ddPCR. Meanwhile, we designed a simple and efficient method for inferring the gene frequency of three blood groups based on ddPCR.
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