Abstract
BackgroundThe characterization of the ABO blood group status is vital for blood transfusion and solid organ transplantation. Several methods for the molecular characterization of the ABO gene, which encodes the alleles that give rise to the different ABO blood groups, have been described. However, the application of those methods has so far been restricted to selected samples and not been applied to population-scale analysis.ResultsWe describe a cost-effective method for high-throughput genotyping of the ABO system by next generation sequencing. Sample specific barcodes and sequencing adaptors are introduced during PCR, rendering the products suitable for direct sequencing on Illumina MiSeq or HiSeq instruments. Complete sequence coverage of exons 6 and 7 enables molecular discrimination of the ABO subgroups and many alleles. The workflow was applied to ABO genotype more than a million samples. We report the allele group frequencies calculated on a subset of more than 110,000 sampled individuals of German origin. Further we discuss the potential of the workflow for high resolution genotyping taking the observed allele group frequencies into account. Finally, sequence analysis revealed 287 distinct so far not described alleles of which the most abundant one was identified in 174 samples.ConclusionsThe described workflow delivers high resolution ABO genotyping at low cost enabling population-scale molecular ABO characterization.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-2687-1) contains supplementary material, which is available to authorized users.
Highlights
The characterization of the ABO blood group status is vital for blood transfusion and solid organ transplantation
Workflow We describe a high-throughput workflow for ABO genotyping based on direct PCR amplicon sequencing on Illumina MiSeq or HiSeq instruments as described for HLA typing in Lange et al [13]
ABO allele frequencies The primary purpose of this project was to supplement HLA genotyping with basic ABO blood group information at low cost
Summary
The characterization of the ABO blood group status is vital for blood transfusion and solid organ transplantation. Several methods for the molecular characterization of the ABO gene, which encodes the alleles that give rise to the different ABO blood groups, have been described. ABO is the clinically most relevant blood group system in transfusion and transplantation medicine [1]. Several medium- to high-throughput molecular typing methods have been developed for the glycosyltransferase encoding ABO gene on human chromosome 9. While these methods generally suffice for clinical applications [7], they do not scale to the requirements of routine upfront ABO genotyping of large cohorts of blood donors [4].
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