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Comparison of DNA extraction methods for human gut microbial community profiling

机译:DNA提取方法对人体肠道微生物群落分析的比较

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摘要

The human gut harbors a vast range of microbes that have significant impact on health and disease. Therefore, gut microbiome profiling holds promise for use in early diagnosis and precision medicine development. Accurate profiling of the highly complex gut microbiome requires DNA extraction methods that provide sufficient coverage of the original community as well as adequate quality and quantity. We tested nine different DNA extraction methods using three commercial kits (TianLong Stool DNA/RNA Extraction Kit (TS), QIAamp DNA Stool Mini Kit (QS), and QIAamp PowerFecal DNA Kit (QP)) with or without additional bead-beating step using manual or automated methods and compared them in terms of DNA extraction ability from human fecal sample. All methods produced DNA in sufficient concentration and quality for use in sequencing, and the samples were clustered according to the DNA extraction method. Inclusion of bead-beating step especially resulted in higher degrees of microbial diversity and had the greatest effect on gut microbiome composition. Among the samples subjected to bead-beating method, TS kit samples were more similar to QP kit samples than QS kit samples. Our results emphasize the importance of mechanical disruption step for a more comprehensive profiling of the human gut microbiome. (C) 2017 The Authors. Published by Elsevier GmbH.
机译:人体肠毛的大量微生物对健康和疾病产生重大影响。因此,肠道微生物组分析占据早期诊断和精密医学发展的承担。高度复杂的肠道微生物组的精确分析需要DNA提取方法,可提供原始社区的充分覆盖以及充足的质量和数量。我们使用三种商用套件测试了九种不同的DNA提取方法(天龙粪便DNA / RNA提取试剂盒(TS),QIAAMP DNA粪便迷你试剂盒(QS)和QIAAMP Powerfecal DNA套件(QP)),使用或无需额外的珠子跳动步骤手动或自动化方法,并以人体粪便样品的DNA提取能力进行比较。所有方法都在足够的浓度和用于测序中使用的DNA,并根据DNA提取方法进行样品。包含珠子跳动步骤,特别是较高的微生物多样性,对肠道微生物组成具有最大的影响。在对珠子跳动法进行的样品中,TS试剂盒样品比QS试剂盒样品更类似于QP试剂盒样品。我们的结果强调了机械中断步骤的重要性,以便更全面的人体肠道微生物曲线探讨。 (c)2017作者。由elsevier GmbH发布。

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