The third-generation sequencing combined with targeted capture technology for high-resolution HLA typing and MHC region haplotype identification
Received date: 2018-11-26
Revised date: 2019-01-25
Online published: 2019-02-25
Supported by
the Young Talents Program of National High-level Personnel of Special Support Program (The “Ten Thousand Talent Program”)
The high-resolution and accurate typing of human leukocyte antigen (HLA) is of great significance for the study of tissue matching in organ transplantation and the correlation between HLA and disease. In this study, the peripheral blood of 12 patients with primary hepatocellular carcinoma was used to compare the advantages and disadvantages of the next- and third-generation sequencing technology for high-resolution HLA typing. In addition, probe capture technology was used to capture the MHC region of YH and HeLa standard cell lines, and a primary hepatocellular carcinoma patient. The captured products were sequenced using PacBio platform to assess the potential of ultra-long reads sequencing technology for analysis of the entire MHC region. Our results showed that: (1) the next- and third-generation sequencing technology can both achieve 6-8 digit high resolution in HLA typing. However, the coverage of the third-generation is significantly better than the next-generation sequencing technology. (2) The ultra-long reads of the third generation sequencing can directly span the entire amplicon region, which has obvious advantages for haplotype phasing, with 92.79% of the HLA genes having accurate phasing results, which is much higher than the 75.65% from the next-generation data. (3) The long-reads from the third generating sequencing can not only be used to assemble the MHC region but also the ability to phase the entire MHC region of 3.6 Mb, thereby helping to clarify the localization information of the mutation sites, alleles and non-coding regions on each MHC haplotype, and providing a theoretical basis for the study of immune and other related diseases.
Jia Chen,Mingyue Shu,Jin Li,Aisi Fu,Fan Yang,Zou Wang,Yirong Li,Zixin Deng,Tiangang Liu . The third-generation sequencing combined with targeted capture technology for high-resolution HLA typing and MHC region haplotype identification[J]. Hereditas(Beijing), 2019 , 41(4) : 337 -348 . DOI: 10.16288/j.yczz.18-282
| [1] | TANG MZ, CAI YL, ZHENG YM, ZENG Y . Association between human leukocyte antigenand nasopharyngeal- carcinoma. Hereditas(Beijing), 2012,34(12):1505-1512. | |||
| [1] | 汤敏中, 蔡永林, 郑裕明, 曾毅 . 人类白细胞抗原与鼻咽癌的相关性. 遗传, 2012,34(12):1505-1512. | |||
| [2] | YANG Zhao-Qing, CHU Jia-You . The research progress of human genetic diversity in China. Hereditas(Beijing), 2012,34(11):1351-1364. | |||
| [2] | 杨昭庆, 褚嘉祐 . 中国人类遗传多样性研究进展. 遗传, 2012,34(11):1351-1364. | |||
| [3] | XU Jun-Pin, DENG Zhi-Hui, JU Gong-Yan, GAO Su-Jing, WANG Da-Meng, HE Liu-Mei, WEI Tian-Chi . Cloning and sequencing HLA-A and -B genomic DNA and analyzing polymorphism in regulatory regions in Chinese Han individuals. Hereditas(Beijing), 2010,32(7):685-693. | |||
| [3] | 徐筠娉, 邓志辉, 邹红岩, 高素青, 王大明, 何柳媚, 魏天莉 . 中国汉族个体HLA-A、-B基因全长序列的测定及调控区多态性. 遗传, 2010,32(7):685-693. | |||
| [4] | Kløverpris HN, Adland E, Koyanagi M, Stryhn A, Harndahl M, Matthews PC, Shapiro R, Walker BD, Ndung'u T, Brander C, Takiguchi M, Buus S, Goulder P . HIV subtype influences HLA-B*07:02-associated HIV disease outcome. AIDS Res Hum Retroviruses, 2014,30(5):468-475. | |||
| [5] | Mallal S, Nolan D, Witt C, Masel G, Martin A, Moore C, Sayer D, Castley A, Mamotte C, Maxwell D, James I, Christiansen FT . Association between presence of HLA-B* 5701, HLA-DR7, and HLA-DQ3 and hypersensitivity to HIV-1 reverse-transcriptase inhibitor abacavir. Lancet, 2002,359(9308):727-732. | |||
| [6] | Galeazzi M, Sebastiani GD, Passiu G, Angelini G, Delfino L, Asherson RA, Khamashta MA, Hughes GR . HLA-DP genotyping in patients with systemic lupus erythematosus: correlations with autoantibody subsets. J Rheumatol, 1992,19(1):42-46. | |||
| [7] | Sasazuki T, Juji T, Morishima Y, Kinukawa N, Kashiwabara H, Inoko H, Yoshida T, Kimura A, Akaza T, Kamikawaji N, Kodera Y, Takaku F . Effect of matching of class I HLA alleles on clinical outcome after transplantation of hematopoietic stem cells from an unrelated donor. Japan Marrow Donor Program. N Engl J Med, 1998,339(17):1177-1185. | |||
| [8] | Donaldson PT, Ho S, Williams R, Johnson PJ . HLA class II alleles in Chinese patients with hepatocellular carcinoma. Liver, 2001,21(2):143-148. | |||
| [9] | Lind C, Ferriola D, Mackiewicz K, Heron S, Rogers M, Slavich L, Walker R, Hsiao T, McLaughlin L, D'Arcy M, Gai X, Goodridge D, Sayer D, Monos D . Next-generation sequencing: the solution for high-resolution, unambiguous human leukocyte antigen typing. Hum Immunol, 2010,71(10):1033-1042. | |||
| [10] | Shiina T, Suzuki S, Ozaki Y, Taira H, Kikkawa E, Shigenari A, Oka A, Umemura T, Joshita S, Takahashi O, Hayashi Y, Paumen M, Katsuyama Y, Mitsunaga S, Ota M, Kulski JK, Inoko H . Super high resolution for single molecule-sequence-based typing of classical HLA loci at the 8-digit level using next generation sequencers. Tiss
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