Being a company with ample experience in whole-gene, high-resolution sequencing, and typing, we have now established a method for blood group sequencing. This method can be used to create a database based on ISBT (International Society of Blood Transfusion) allele information.
The term “blood group” refers to the entire blood group system comprising red blood cell (RBC) antigens whose specificity is controlled by a series of genes that can be allelic or linked very closely on the same chromosome. Currently, there are 43 blood group systems described by the ISBT. Their typing is almost exclusively based on serology or assays detecting a limited number of SNPs. Sequencing of rare blood groups, especially whole-gene, is rare and there is currently no comprehensive database with systematic nomenclature to keep track of all variants. With the method we established, data can be generated and novel sequences can be contributed to a curated database, and become accessible to the scientific community.
Method
We designed amplification primers for 7 blood group systems; Kell (KEL), Lewis (LE), Duffy (FY), Diego DI), Landsteiner-Wiener (LW), H (H), and Vel (VEL). For the sequencing, we used Illumina MiSeq 2×150 and MinION R9.4. Amplicon length ranged from 2 to 22 kb. Phasing regions are far lower in MinION sequencing. To present, we have identified a large number of variants from only 7 samples (14 alleles).
Whole gene sequencing shows both coding and non-coding variants. MiSeq and ONT can be combined for enhanced phasing. PacBio sequencing combines features of MiSeq and ONT and we are confident that it will enhance phasing and therefore be favorable for blood group whole gene sequencing.
A proper understanding of the blood group system, their clinical significance, typing and cross-matching tests, and current perspective are very important to prevent transfusion-related complications. Nonetheless, the knowledge on the blood group system is necessary to approach blood group-linked diseases which are still at the stage of research. Additional sample testing is also needed to indicate the scale of genetic variability.
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