Division of Gastroenterology and Hepatology, Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan; Department of Internal Medicine, National Taiwan University College of Medicine, Taipei, Taiwan; Graduate Institute of Clinical Medicine, National Taiwan University College of Medicine, Taipei, Taiwan.
Department of Internal Medicine, National Taiwan University Hospital Bei-Hu Branch, Taipei, Taiwan; Institute of Food Science and Technology, National Taiwan University, Taipei, Taiwan.
J Formos Med Assoc. 2020 Dec;119(12):1791-1798. doi: 10.1016/j.jfma.2020.01.013. Epub 2020 Feb 26.
BACKGROUND/PURPOSE: Appropriate storage of fecal samples is a critical step for the unbiased analysis of microbial communities in metagenomic studies. Rapid freezing at -80 °C is usually considered to be best practice, but this approach is challenging. DNA stabilizing kits may provide a more convenient method to preserve and store clinical samples. We evaluated the reliability of two collection kits (Stratec stool collection tube with stabilizer, #1038111200 and OMNIgene.GUT OMR-200) on preserving fecal microbiota. METHODS: Samples were collected from two locations of the fecal specimen, in four healthy volunteers. The samples were sub-aliquoted and stored in a -80 °C freezer, in Stratec and OMNIgene.GUT (incubation at ambient temperature for 0, 3, or 7 days). The fecal microbial composition was assessed by 16S rRNA sequencing. RESULTS: We found that alpha diversity was not significantly affected by storage conditions. Samples stored in DNA stabilizers were still representative of the original microbial community after 7 days at ambient temperature. Individual differences were found to have a greater contribution to the differences in microbial community composition than storage conditions or sampling location. Samples subjected to stabilizers displayed microbial community shifts compared with immediately frozen samples. A linear discriminant analysis effect size (LEfSe) analysis showed that the relative abundances of Faecalibacterium were significantly higher in samples stored in Stratec kits. CONCLUSION: Our study reveals that both Stratec and OMNIgene.GUT kits provide good microbiome preservation for up to 7 days in ambient temperature and would represent good options for fecal sample collection in large scale, population-based studies.
背景/目的:适当储存粪便样本是对宏基因组研究中微生物群落进行无偏分析的关键步骤。通常认为 -80°C 的快速冷冻是最佳实践,但这种方法具有挑战性。DNA 稳定剂试剂盒可能提供一种更方便的方法来保存和储存临床样本。我们评估了两种采集试剂盒(Stratec 粪便采集管与稳定剂,#1038111200 和 OMNIgene.GUT OMR-200)在保存粪便微生物群方面的可靠性。
方法:从四个健康志愿者粪便样本的两个部位采集样本。将样本分小份并储存在-80°C 冰箱中,在 Stratec 和 OMNIgene.GUT 中(在环境温度下孵育 0、3 或 7 天)。通过 16S rRNA 测序评估粪便微生物组成。
结果:我们发现,储存条件对 alpha 多样性没有显著影响。在环境温度下储存 7 天后,储存在 DNA 稳定剂中的样本仍然代表原始微生物群落。个体差异对微生物群落组成差异的贡献大于储存条件或采样位置。与立即冷冻的样本相比,经稳定剂处理的样本显示出微生物群落的变化。线性判别分析效应量(LEfSe)分析显示,Stratec 试剂盒中储存的粪杆菌相对丰度显著更高。
结论:我们的研究表明,Stratec 和 OMNIgene.GUT 试剂盒在环境温度下储存长达 7 天都能很好地保存微生物组,并且对于基于人群的大规模粪便样本采集来说是很好的选择。
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