Distributions of MICA and MICB Alleles Typed by Amplicon‐Based Next‐Generation Sequencing in South Koreans.

Major histocompatibility complex class I chain‐related genes A and B (MICA and MICB) play a role as ligands in activating the NKG2D receptor expressed in natural killer cells, γδ T‐cells and αβ CD8 T‐cells and have been defined in human diseases and haematopoietic stem cell transplantation (HSCT). MICA and MICB alleles were genotyped at the three‐field level by amplicon‐based next‐generation sequencing (NGS) using a MiSeqDx system and compared with the results from previous studies in healthy South Korean donors. Exons 2–5 of MICA and exons 2–4 of MICB were amplified using a multiplex polymerase chain reaction (PCR). Sequence reads of ≥ 51 depth counts were consistently obtained for each sample exon, and target exons were determined to match reference sequences contained in the IPD‐IMGT/HLA database. MICA and MICB alleles were tested using exon combinations. The program was designed to recognise specific sequences and discriminate between the MICA*008:01:01/*027 alleles. A total of 22 alleles were found in MICA and MICB. We observed 1 HLA‐C ~ HLA‐B ~ MICA ~ MICB ~ HLA‐DRB1 haplotype with significant linkage disequilibrium between alleles at all neighbouring HLA loci. These results are consistent with previous microarray results. Genotyping of MICA and MICB was possible using 11‐loci HLA genes. We updated the distribution of MICA and MICB based on three‐field allele and haplotype frequencies containing linkage disequilibrium in South Koreans using amplicon‐based NGS. These data suggest that high‐resolution MICA and MICB typing data obtained using NGS may aid in performing HSCT and disease association studies. [ABSTRACT FROM AUTHOR]