BSP-01: Full Four-Digit Typing for Class I and II HLA Genes

The Basic Science Program will receive genomic DNA at a concentration of 50 ng/ul. Human leukocyte antigen (HLA) typing will be performed using a targeted next-generation sequencing (NGS) method. Briefly, locus-specific primers are used to amplify a total of 23 polymorphic exons of HLA-A, B, and C (exons 1 to 4); DPA1 (exon 2); DPB1 (exons 2 and 3); DQA1 (exon 2), DQB1 (exons 2 and 3); DRB1 (exons 2 and 3); and DRB3, 4, and 5 (exon 2) genes with Fluidigm Access Array (Fluidigm Singapore PTE Ltd, Singapore). The 23 Fluidigm polymerase chain reaction (PCR) amplicons are pooled and subjected to sequencing on an illumina MiSeq sequencer (Illumina, San Diego, CA 92122 USA). The Basic Science Program will analyze the results and assign HLA genotypes using the Omixon HLA Explore software (Omixon, Budapest, Hungary). Alleles for all the above-mentioned HLA genes will be reported at the four-digit resolution level to customers. Haplotypes will also be assessed. Sufficient basic consultation with a subject matter expert is included with the service.