Shotgun Metagenomic analysis of cutaneous microbiome in patients with atopic dermatitis

M. Ramadan, S. Solyman, M. Yones, Hamada Halaby, Yaser Abdalla, Amr Hanora
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引用次数: 2

Abstract

Atopic dermatitis (AD) is a chronic complex inflammatory skin disorder. AD characterized by multiple contributing factors, such as impairment in skin barrier integrity, participation of either innate or adaptive immunity arms, in addition to the pivotal roles of cutaneous microbiome. The analysis of cutaneous microbiota was performed using shotgun metagenomic sequencing on an Illumina HiSeq platform in parallel with 16s rRNA genes using MiSeq sequencing. Two samples for each sequencing technique were collected from one child and one adult subjects with AD. Metagenomic data from 16srRNA was in accordance with those from shotgun sequencing. Taxonomic profile of samples at genus level showed overrepresentation of Streptococcus in child and Staphylococcus in adults. Species level analysis of reads revealed monoclonality of Staphylococcus aureus community in comparison to heterogeneity of Staphylococcus epidermidis. 16S rRNA gene based analysis was unable to provide comprehensive description of bacterial communities, while shotgun metagenomic analyses described the composition of microbiota with high resolution at different taxonomic levels especially subspecies and single nucleotide variants levels. Finally, further studies using shotgun metagenomic approaches will be required for characterization of microbiome in healthy and diseased subjects.
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特应性皮炎患者皮肤微生物群的散弹枪宏基因组分析
特应性皮炎(AD)是一种慢性复杂炎症性皮肤病。除了皮肤微生物群的关键作用外,AD的特征还包括多种因素,如皮肤屏障完整性受损、先天或适应性免疫臂的参与。在Illumina HiSeq平台上使用散弹枪宏基因组测序对皮肤微生物群进行分析,同时使用MiSeq测序对16s rRNA基因进行分析。每种测序技术分别从一名AD儿童和一名AD成人受试者中采集两份样本。16srRNA的宏基因组数据与霰弹枪测序结果一致。属水平样本的分类分析显示,儿童链球菌和成人葡萄球菌的代表性过高。reads的物种水平分析显示金黄色葡萄球菌群落单克隆,而表皮葡萄球菌的异质性。基于16S rRNA基因的分析无法提供细菌群落的全面描述,而霰弹枪宏基因组分析在不同的分类水平特别是亚种和单核苷酸变异水平上能够高分辨率地描述微生物群的组成。最后,需要使用霰弹枪宏基因组方法进行进一步的研究,以表征健康和患病受试者的微生物组。
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