Al Kait F, Bisanz Jordan E, Gloor Gregory B, Reid Gregor, Burton Jeremy P
Departments of Microbiology & Immunology, University of Western Ontario, London, Ontario, Canada; Canadian Centre for Human Microbiome and Probiotics, London, Ontario, Canada; Lawson Health Research Institute, London, Ontario, Canada.
Canadian Centre for Human Microbiome and Probiotics, London, Ontario, Canada; Department of Biochemistry, University of Western Ontario, London, Ontario, Canada.
J Microbiol Methods. 2018 Jan;144:117-121. doi: 10.1016/j.mimet.2017.11.014. Epub 2017 Nov 16.
The increasing interest on the impact of the gut microbiota on health and disease has resulted in multiple human microbiome-related studies emerging. However, multiple sampling methods are being used, making cross-comparison of results difficult. To avoid additional clinic visits and increase patient recruitment to these studies, there is the potential to utilize at-home stool sampling. The aim of this pilot study was to compare simple self-sampling collection and storage methods.
To simulate storage conditions, stool samples from three volunteers were freshly collected, placed on toilet tissue, and stored at four temperatures (-80, 7, 22 and 37°C), either dry or in the presence of a stabilization agent (RNAlater®) for 3 or 7days. Using 16S rRNA gene sequencing by Illumina, the effect of storage variations for each sample was compared to a reference community from fresh, unstored counterparts. Fastq files may be accessed in the NCBI Sequence Read Archive: Bioproject ID PRJNA418287.
Microbial diversity and composition were not significantly altered by any storage method. Samples were always separable based on participant, regardless of storage method suggesting there was no need for sample preservation by a stabilization agent.
In summary, if immediate sample processing is not feasible, short term storage of unpreserved stool samples on toilet paper offers a reliable way to assess the microbiota composition by 16S rRNA gene sequencing.
对肠道微生物群对健康和疾病影响的兴趣日益增加，导致多项与人类微生物组相关的研究不断涌现。然而，目前使用的采样方法多种多样，使得结果的交叉比较变得困难。为了避免额外的门诊就诊并增加参与这些研究的患者数量，利用家庭粪便采样具有可行性。本试点研究的目的是比较简单的自我采样收集和存储方法。
为模拟存储条件，从三名志愿者新鲜采集粪便样本，置于卫生纸 上，分别在四种温度（-80、7、22 和 37°C）下干燥或在存在稳定试剂（RNAlater®）的情况下存储 3 天或 7 天。通过 Illumina 进行 16S rRNA 基因测序，将每个样本存储变化的影响与来自新鲜、未存储样本的参考群落进行比较。Fastq 文件可在 NCBI 序列读取存档中获取：生物项目 ID PRJNA418287。
任何存储方法均未显著改变微生物多样性和组成。无论存储方法如何，样本始终可根据参与者进行区分，这表明无需使用稳定试剂保存样本。
总之，如果无法立即进行样本处理，将未保存的粪便样本短期存储在卫生纸上，为通过 16S rRNA 基因测序评估微生物群组成提供了一种可靠的方法。
