Center for Applied Microbiome Science, Pathogen and Microbiome Institute, Northern Arizona University, Flagstaff, Arizona, USA.
Biodesign Swette Center for Environmental Biotechnology, Arizona State University, Tempe, Arizona, USA.
mSphere. 2019 Apr 10;4(2):e00208-19. doi: 10.1128/mSphere.00208-19.
Studies of the intestinal microbiome commonly utilize stool samples to measure the microbial composition in the distal gut. However, collection of stool can be difficult from some subjects under certain experimental conditions. Sampling of fecal material using sterile swabs can streamline sample collection, handling, and processing. In this study, we validate the use of swabs of fecal matter to approximate measurements of microbiota in stool using 16S rRNA gene Illumina amplicon sequencing and evaluate the effects of shipping time at ambient temperatures on accuracy. The results indicate that swab samples reliably replicate the stool microbiota bacterial composition, alpha diversity, and beta diversity when swabs are processed quickly (≤2 days) but that sample quality quickly degrades after this period and is accompanied by increased abundances of Fresh swabs appear to be a viable alternative to stool sampling when standard collection methods are challenging, but extended exposure to ambient temperatures prior to processing threatens sample integrity. Collection of fecal swab samples simplifies handling, processing, and archiving compared to collection of stool. This study confirms that fecal swabs reliably replicate the bacterial composition and diversity of stool samples, provided that the swabs are processed shortly after collection. These findings support the use of fecal swabs, when shipping and handling are done properly, to streamline measurements of intestinal microbiota.
研究肠道微生物组通常使用粪便样本来测量远端肠道中的微生物组成。然而,在某些实验条件下,从某些研究对象中收集粪便可能很困难。使用无菌拭子采集粪便样本可以简化样本采集、处理和加工。在这项研究中,我们使用 16S rRNA 基因 Illumina 扩增子测序来验证使用拭子粪便来近似测量粪便中微生物群的方法,并评估在环境温度下运输时间对准确性的影响。结果表明,当拭子在短时间内(≤2 天)处理时,拭子样本可靠地复制了粪便微生物群的细菌组成、α多样性和β多样性,但在此期间之后,样本质量迅速下降,并且伴随着新鲜拭子的增加丰度似乎是在标准收集方法具有挑战性时替代粪便采样的可行选择,但在处理之前长时间暴露于环境温度会威胁到样本完整性。与收集粪便相比,收集粪便拭子可简化处理、处理和存档。本研究证实,只要在收集后不久进行处理,粪便拭子就可以可靠地复制粪便样本的细菌组成和多样性。这些发现支持在运输和处理得当的情况下使用粪便拭子来简化肠道微生物组的测量。