https://researchopenworld.com/avoid-using-sample-with-more-than-70-blasts-for-hla-typing-to-reduce-erroneous-homozygous-hla-typing-results-by-microbead-assay/#

作者：Jeong-Shi Lin, Li-Hsuan Lee, Hsueng‐Mei Liu, Ying‐Ju Chen · 发表于：https://researchopenworld.com/category/journal-of-molecular-genetics/ · 年份：2020 · DOI：10.31038/jmg.2020312

Background: Loss of heterozygosity (LOH) at the human leukocyte antigen (HLA) region could lead to erroneous homozygous HLA typing results. Materials and methods: We investigated HLA typing on peripheral blood samples derived from a patient with acute myeloid leukemia (AML) at diagnosis and remission by Luminex microbead assay. LOH was analyzed by short tandem repeat (STR) analysis at markers of long arm and short arm of chromosome 6 and single-nucleotide polymorphism (SNP) array analysis. For DNA mixing test to define the detection threshold for heterozygous HLA genotypes, we selected 6 samples of homozygous HLA typing and 6 samples of heterozygous HLA typing. Results: At diagnosis (blasts 77% in peripheral blood), HLA typing revealed A*11, B*15:02/88(B75), C*08:01/08, DRB1*08, DQB1*06. Short tandem repeat (STR) analysis of peripheral blood revealed segmental uniparental disomy (UPD) of chromosome 6 (LOH of marker at 6p22, but no LOH at other markers of short arm and long arm of chromosome 6). Analysis of LOH by single-nucleotide polymorphism (SNP) array analysis demonstrated no copy number variations, but LOH on chromosome 6 [34.6Mb] and on chromosome 13 [92.8Mb]. At remission, HLA typing was A*11, A*24, B*15:02/112(B75), B*40:01/22N (B60), C*07:02/32N, C*08:01/08, DRB1*08, DRB1*14, DQB1*05, DQB1*06. DNA mixing experiments revealed that the minimum threshold for detecting HLA heterozygosity using Luminex technology is < 70% homozygous sample. Conclusion: We describe an erro...