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Single molecule real‐time DNA sequencing of the full HLA‐E gene for 212 reference cell lines
Author(s) -
Lucas Jonathan A. M.,
Hayhurst James D.,
Turner Thomas R.,
Gymer Arthur W.,
Leen Gayle,
Robinson James,
Marsh Steven G. E.,
Mayor Neema P.
Publication year - 2020
Publication title -
hla
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.347
H-Index - 99
eISSN - 2059-2310
pISSN - 2059-2302
DOI - 10.1111/tan.13882
Subject(s) - genotyping , human leukocyte antigen , zygosity , genotype , genetics , haplotype , biology , allele , microbiology and biotechnology , gene , dna sequencing , dna , antigen
We have developed a genotyping assay that produces fully phased, unambiguous HLA‐E genotyping using Pacific Biosciences' single molecule real‐time DNA sequencing. In total 212 cell lines were genotyped, including the panel of 107 established at the 10th International Histocompatibility Workshop. Our results matched the previously known HLA‐E genotype in 94 (44.3%) cell lines, in all cases either improving or equalling previous genotyping resolution. Three (1.4%) cells had discrepant HLA‐E genotyping data and 115 (54.2%) had no previous HLA‐E data. The HLA‐E genotypes for four (1.9%) cell lines resulted in a change of zygosity by identifying two distinct haplotypes. We discovered eight novel HLA‐E alleles, extended the known reference sequence of seven and confirmed the existence of a further 10.