Next‐generation sequencing: academic overkill or high‐resolution routine blood group genotyping?

Abstract

Blood group genotyping (BGG) has been in routine clinical practice ever since the molecular determination of blood groups was achieved, during the early to mid 1990s. These early methods were dependent on allele‐specific PCR to detect simple single nucleotide polymorphisms (SNPs) responsible for blood group expression. As our knowledge regarding the molecular background of blood groups advanced, so did the numbers of SNPs requiring detection which necessitated the switch from allele‐specific PCR to array‐based technology, notably slide and bead‐based approaches. All these methods thus described are totally dependent on the predefined genetic basis of each SNP. Hybrid blood group genes (as found in RH, ABO and MNS systems) are difficult to define by all of these aforementioned techniques. With the arrival of cheap next‐generation sequencing (NGS) approaches in the past 5 years, we have conducted long‐range PCR (LR‐PCR) coupled with NGS determination of the major blood group genes. By analysis of these data using an Ion Torrent Personal Genome Machine™ (PGM™), it is readily apparent that NGS can highly effectively be applied with high resolution to blood grouping at costs no more than current array‐based platforms. However, the complexity of the data obtained need careful filtering for effective clinical utilization, but provides useful insight on the evolution of blood groups and their environmental impacts which will be of undoubted value as an academic exercise, but of minimal cost (if any) to the original testing.