title: An integrated tool to study MHC region: accurate SNV detection and HLA genes typing in human MHC region using targeted high-throughput sequencing.
abstract: The major histocompatibility complex (MHC) is one of the most variable and gene-dense regions of the human genome. Most studies of the MHC, and associated regions, focus on minor variants and HLA typing, many of which have been demonstrated to be associated with human disease susceptibility and metabolic pathways. However, the detection of variants in the MHC region, and diagnostic HLA typing, still lacks a coherent, standardized, cost effective and high coverage protocol of clinical quality and reliability. In this paper, we presented such a method for the accurate detection of minor variants and HLA types in the human MHC region, using high-throughput, high-coverage sequencing of target regions. A probe set was designed to template upon the 8 annotated human MHC haplotypes, and to encompass the 5 megabases (Mb) of the extended MHC region. We deployed our probes upon three, genetically diverse human samples for probe set evaluation, and sequencing data show that ∼97% of the MHC region, and over 99% of the genes in MHC region, are covered with sufficient depth and good evenness. 98% of genotypes called by this capture sequencing prove consistent with established HapMap genotypes. We have concurrently developed a one-step pipeline for calling any HLA type referenced in the IMGT/HLA database from this target capture sequencing data, which shows over 96% typing accuracy when deployed at 4 digital resolution. This cost-effective and highly accurate approach for variant detection and HLA typing in the MHC region may lend further insight into immune-mediated diseases studies, and may find clinical utility in transplantation medicine research. This one-step pipeline is released for general evaluation and use by the scientific community.
authors.affiliation: Science and Technology Department, BGI-Shenzhen, Shenzhen, China ; Department of Biology, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Hongzhi
authors.initials: H
authors.lastname: Cao
authors.forename: Jinghua
authors.initials: J
authors.lastname: Wu
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.forename: Hui
authors.initials: H
authors.lastname: Jiang
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.forename: Xiao
authors.initials: X
authors.lastname: Liu
authors.forename: Yinyin
authors.initials: Y
authors.lastname: Xu
authors.forename: Dequan
authors.initials: D
authors.lastname: Liang
authors.forename: Peng
authors.initials: P
authors.lastname: Gao
authors.forename: Yepeng
authors.initials: Y
authors.lastname: Sun
authors.forename: Benjamin
authors.initials: B
authors.lastname: Gifford
authors.forename: Mark
authors.initials: M
authors.lastname: D'Ascenzo
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Liu
authors.forename: Laurent C A M
authors.initials: LC
authors.lastname: Tellier
authors.forename: Fang
authors.initials: F
authors.lastname: Yang
authors.forename: Xin
authors.initials: X
authors.lastname: Tong
authors.forename: Dan
authors.initials: D
authors.lastname: Chen
authors.forename: Jing
authors.initials: J
authors.lastname: Zheng
authors.forename: Weiyang
authors.initials: W
authors.lastname: Li
authors.forename: Todd
authors.initials: T
authors.lastname: Richmond
authors.forename: Xun
authors.initials: X
authors.lastname: Xu
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.forename: Yingrui
authors.initials: Y
authors.lastname: Li
mesh_terms: D005838:Genotype; D006239:Haplotypes / Q000235:genetics; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006650:Histocompatibility Testing; D006801:Humans; D008285:Major Histocompatibility Complex / Q000235:genetics*
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