Research Group of Gut Microbiome, Division of Nutrition and Metabolism Research, Korea Food Research Institute, Jeollabuk-do 55365, Republic of Korea.
Research Group of Gut Microbiome, Division of Nutrition and Metabolism Research, Korea Food Research Institute, Jeollabuk-do 55365, Republic of Korea; Department of Food Biotechnology, Korea University of Science and Technology, Daejeon 34113, Republic of Korea.
Syst Appl Microbiol. 2018 Mar;41(2):151-157. doi: 10.1016/j.syapm.2017.11.008. Epub 2017 Dec 21.
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.
人类肠道中栖息着大量的微生物,这些微生物对健康和疾病有着重要的影响。因此,肠道微生物组分析有望用于早期诊断和精准医学的发展。要准确分析高度复杂的肠道微生物组,需要使用能够充分覆盖原始群落、具有足够质量和数量的 DNA 提取方法。我们使用三种商业试剂盒(TianLong Stool DNA/RNA 提取试剂盒(TS)、QIAamp DNA Stool Mini 试剂盒(QS)和 QIAamp PowerFecal DNA 试剂盒(QP)),分别或同时使用手动或自动方法,测试了九种不同的 DNA 提取方法,比较了它们从人粪便样本中提取 DNA 的能力。所有方法都能提取出足够浓度和质量的 DNA 用于测序,并且根据 DNA 提取方法对样本进行了聚类。加入珠磨步骤尤其会增加微生物多样性的程度,并对肠道微生物组的组成产生最大的影响。在进行珠磨法的样本中,TS 试剂盒样本比 QS 试剂盒样本更类似于 QP 试剂盒样本。我们的结果强调了机械破碎步骤对于更全面地分析人类肠道微生物组的重要性。