Next-generation sequencing for HLA typing of class I loci.

Next-generation sequencing for HLA typing of class I loci.
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I级基因座HLA键入的下一代测序。

DOI:
10.1186/1471-2164-12-42
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发表时间:
2011-01-18
期刊:
影响因子:
4.4
通讯作者:
de Bakker PI
de Bakker PI
中科院分区:
生物学2区
文献类型:
--
作者:
Erlich RL;Jia X;Anderson S;Banks E;Gao X;Carrington M;Gupta N;DePristo MA;Henn MR;Lennon NJ;de Bakker PI

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跨MHC的全面序列特征对于成功的器官移植和遗传关联研究是重要的。为此,我们开发了一种自动化的样本制备、分子条形码和多路复用协议,用于扩增和测定I类人类白细胞抗原基因座的序列。我们将这一过程与一种新的HLA调用算法相结合,以确定每个基因座上最可能的等位基因对。我们已经以来自全球四个群体的270名HapMap个体为基准对我们的协议进行了基准测试,在4位数分辨率下,准确率为96.4%。对95个HapMap个体进行了4位数分辨率下98.6%的准确率测试,该方法更适合于大样本量,其中在PCR过程中添加分子条形码而不是文库构建。在454FLX钛平台上进行的下一代测序是一种可靠、高效和可扩展的人类白细胞抗原分型技术。
Comprehensive sequence characterization across the MHC is important for successful organ transplantation and genetic association studies. To this end, we have developed an automated sample preparation, molecular barcoding and multiplexing protocol for the amplification and sequence-determination of class I HLA loci. We have coupled this process to a novel HLA calling algorithm to determine the most likely pair of alleles at each locus. We have benchmarked our protocol with 270 HapMap individuals from four worldwide populations with 96.4% accuracy at 4-digit resolution. A variation of this initial protocol, more suitable for large sample sizes, in which molecular barcodes are added during PCR rather than library construction, was tested on 95 HapMap individuals with 98.6% accuracy at 4-digit resolution. Next-generation sequencing on the 454 FLX Titanium platform is a reliable, efficient, and scalable technology for HLA typing.
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