Authors:
B. Matern (Maastricht, NL, NL)
T. Slangen (Maastricht, NL)
M. Groeneweg (Maastricht, NL)
M. Tilanus (Maastricht, NL)
C. Voorter (Maastricht, NL)
Typing of the ABO blood groups, which encodes the transferase that catalyzes the last step in the synthesis of the A, B, and H antigens, is a prerequisite for both the recipient and donor in solid organ transplantation, stem cell transplantation, and blood transfusion. Historically, blood group typing has typically been done with serology, but in a number of cases, such as registry typing with buccal swabs and samples from cord blood, only DNA is available. The blood group types A, B, and O show clear differences in the genetic sequence of the ABO gene, and therefore DNA sequencing offers a valuable alternative to serology in these cases. In this study we have used MinION Nanopore sequencing to obtain full length sequences of the ABO gene. Two long-range overlapping amplicons were prepared which span the complete 18kb length of the ABO gene for a set of samples which represent the six possible ABO combinations (AA, BB, AO, BO, AB and OO). The amplicons were sequenced on the Oxford Nanopore MinION, and reads were aligned with known reference sequences obtained from the NCBI dbRBC database. DNA sequence data was sufficient to differentiate between the six possible ABO combinations based on 3 polymorphic positions. Although the current analysis focuses on the polymorphism that defines the blood group types as a proof of principle, the acquired full-length sequences can provide information about the extensiveness of polymorphism present in the ABO alleles, and help complete the full-length ABO allele database. Overall, this study shows that Nanopore sequencing on the MinION represents a novel platform for full-length high-throughput sequencing of ABO genes, suitable for cases where only DNA is available.