Meisel Jacquelyn S, Hannigan Geoffrey D, Tyldsley Amanda S, SanMiguel Adam J, Hodkinson Brendan P, Zheng Qi, Grice Elizabeth A
Department of Dermatology, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, USA.
Department of Dermatology, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, USA.
J Invest Dermatol. 2016 May;136(5):947-956. doi: 10.1016/j.jid.2016.01.016. Epub 2016 Jan 29.
Culture-independent studies to characterize skin microbiota are increasingly common, due in part to affordable and accessible sequencing and analysis platforms. Compared to culture-based techniques, DNA sequencing of the bacterial 16S ribosomal RNA (rRNA) gene or whole metagenome shotgun (WMS) sequencing provides more precise microbial community characterizations. Most widely used protocols were developed to characterize microbiota of other habitats (i.e., gastrointestinal) and have not been systematically compared for their utility in skin microbiome surveys. Here we establish a resource for the cutaneous research community to guide experimental design in characterizing skin microbiota. We compare two widely sequenced regions of the 16S rRNA gene to WMS sequencing for recapitulating skin microbiome community composition, diversity, and genetic functional enrichment. We show that WMS sequencing most accurately recapitulates microbial communities, but sequencing of hypervariable regions 1-3 of the 16S rRNA gene provides highly similar results. Sequencing of hypervariable region 4 poorly captures skin commensal microbiota, especially Propionibacterium. WMS sequencing, which is resource and cost intensive, provides evidence of a community's functional potential; however, metagenome predictions based on 16S rRNA sequence tags closely approximate WMS genetic functional profiles. This study highlights the importance of experimental design for downstream results in skin microbiome surveys.
由于测序和分析平台价格亲民且易于使用,越来越多不依赖培养的研究用于表征皮肤微生物群。与基于培养的技术相比,细菌16S核糖体RNA(rRNA)基因的DNA测序或全宏基因组鸟枪法(WMS)测序能更精确地表征微生物群落。大多数广泛使用的方案是为表征其他栖息地(如胃肠道)的微生物群而开发的,尚未对其在皮肤微生物组调查中的效用进行系统比较。在此,我们为皮肤研究界建立了一个资源,以指导表征皮肤微生物群的实验设计。我们将16S rRNA基因的两个广泛测序区域与WMS测序进行比较,以概括皮肤微生物组群落组成、多样性和基因功能富集情况。我们发现WMS测序最能准确概括微生物群落,但16S rRNA基因高变区1-3的测序提供了高度相似的结果。高变区4的测序很难捕获皮肤共生微生物群,尤其是丙酸杆菌。WMS测序资源和成本密集,提供了群落功能潜力的证据;然而,基于16S rRNA序列标签的宏基因组预测与WMS基因功能概况非常接近。这项研究强调了实验设计对皮肤微生物组调查下游结果的重要性。