ImmunoGlobulin galaxy (IGGalaxy) for simple determination and quantitation of immunoglobulin heavy chain rearrangements from NGS.

ImmunoGlobulin galaxy (IGGalaxy) for simple determination and quantitation of immunoglobulin heavy chain rearrangements from NGS.
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DOI:
10.1186/s12865-014-0059-7
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发表时间:
2014-12-13
期刊:
影响因子:
3
通讯作者:
Stubbs AP
Stubbs AP
中科院分区:
医学4区
文献类型:
--
作者:
Moorhouse MJ;van Zessen D;IJspeert H;Hiltemann S;Horsman S;van der Spek PJ;van der Burg M;Stubbs AP

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免疫球蛋白重链 (IGH) 基因重排的序列分析和频率分析是研究健康和疾病中的免疫组库、免疫反应和免疫失调的有力工具。面临的挑战是为使用 NGS 技术确定 VDJ 库的小型和大型实验室提供用户友好、安全和可重复的分析服务。在这项研究中,我们描述了免疫球蛋白 Galaxy (IGGalaxy)——一种基于网络的便捷应用程序,用于分析下一代测序结果并报告 IGH 基因重排,以进行谱库和克隆性研究。 IGGalaxy 有两种分析选项,一种使用内置的 igBLAST 算法,第二种使用 IMGT 的输出;无论哪种情况,都可以获得所测试的 B 细胞群的指令集摘要。 IGGalaxy 支持 igBLAST 和 IMGT/HIGHV-QUEST 的多样本和多重复输入分析。我们使用标准数据集 S22 证明了该平台的技术有效性,该数据集用于对抗体比对实用程序的性能进行基准测试,与之前的结果具有 99.9% 的一致性。对 RAG 缺陷患者样本的 NGS 数据进行重新分析证明了该工具的有效性和用户友好性。 IGGalaxy 为临床研究人员提供了对每个测序个体的 B 细胞群以及对照基因组和致病基因组之间的详细了解。 IGGalaxy 专为 454 个 NGS 结果而开发,但能够分析替代 NGS 数据(例如 Illumina、Ion Torrent)。我们演示了如何使用 Galaxy 虚拟机来确定参考数据的 VDJ 指令集以及取自免疫缺陷患者的 B 细胞。 IGGalaxy 可作为虚拟机下载并在桌面 PC 或服务器上使用。本文的在线版本 (doi:10.1186/s12865-014-0059-7) 包含补充材料,可供授权用户使用。
Sequence analysis of immunoglobulin heavy chain (IGH) gene rearrangements and frequency analysis is a powerful tool for studying the immune repertoire, immune responses and immune dysregulation in health and disease. The challenge is to provide user friendly, secure and reproducible analytical services that are available for both small and large laboratories which are determining VDJ repertoire using NGS technology. In this study we describe ImmunoGlobulin Galaxy (IGGalaxy)- a convenient web based application for analyzing next-generation sequencing results and reporting IGH gene rearrangements for both repertoire and clonality studies. IGGalaxy has two analysis options one using the built in igBLAST algorithm and the second using output from IMGT; in either case repertoire summaries for the B-cell populations tested are available. IGGalaxy supports multi-sample and multi-replicate input analysis for both igBLAST and IMGT/HIGHV-QUEST. We demonstrate the technical validity of this platform using a standard dataset, S22, used for benchmarking the performance of antibody alignment utilities with a 99.9 % concordance with previous results. Re-analysis of NGS data from our samples of RAG-deficient patients demonstrated the validity and user friendliness of this tool. IGGalaxy provides clinical researchers with detailed insight into the repertoire of the B-cell population per individual sequenced and between control and pathogenic genomes. IGGalaxy was developed for 454 NGS results but is capable of analyzing alternative NGS data (e.g. Illumina, Ion Torrent). We demonstrate the use of a Galaxy virtual machine to determine the VDJ repertoire for reference data and from B-cells taken from immune deficient patients. IGGalaxy is available as a VM for download and use on a desktop PC or on a server. The online version of this article (doi:10.1186/s12865-014-0059-7) contains supplementary material, which is available to authorized users.
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发表时间: 2005-10-01
期刊: GENOME RESEARCH
影响因子: 7
作者:
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发表时间: 2010-01
影响因子: --
作者:
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期刊: Genome biology
影响因子: 12.3
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