Widjaja Frances, Rietjens Ivonne M C M
Division of Toxicology, Wageningen University & Research, 6708 WE Wageningen, The Netherlands.
Biomedicines. 2023 Sep 28;11(10):2658. doi: 10.3390/biomedicines11102658.
The composition, viability and metabolic functionality of intestinal microbiota play an important role in human health and disease. Studies on intestinal microbiota are often based on fecal samples, because these can be sampled in a non-invasive way, although procedures for sampling, processing and storage vary. This review presents factors to consider when developing an automated protocol for sampling, processing and storing fecal samples: donor inclusion criteria, urine-feces separation in smart toilets, homogenization, aliquoting, usage or type of buffer to dissolve and store fecal material, temperature and time for processing and storage and quality control. The lack of standardization and low-throughput of state-of-the-art fecal collection procedures promote a more automated protocol. Based on this review, an automated protocol is proposed. Fecal samples should be collected and immediately processed under anaerobic conditions at either room temperature (RT) for a maximum of 4 h or at 4 °C for no more than 24 h. Upon homogenization, preferably in the absence of added solvent to allow addition of a buffer of choice at a later stage, aliquots obtained should be stored at either -20 °C for up to a few months or -80 °C for a longer period-up to 2 years. Protocols for quality control should characterize microbial composition and viability as well as metabolic functionality.
肠道微生物群的组成、活力和代谢功能在人类健康与疾病中起着重要作用。对肠道微生物群的研究通常基于粪便样本，因为这些样本可以通过非侵入性方式采集，尽管采样、处理和储存程序各不相同。本综述介绍了在制定粪便样本采样、处理和储存的自动化方案时需要考虑的因素：供体纳入标准、智能马桶中的尿便分离、匀浆、分装、溶解和储存粪便材料所用缓冲液的用途或类型、处理和储存的温度及时间以及质量控制。现有粪便采集程序缺乏标准化且通量低，因此需要一个更自动化的方案。基于本综述，提出了一个自动化方案。粪便样本应在厌氧条件下立即处理，在室温（RT）下最多处理4小时，或在4°C下处理不超过24小时。匀浆时，最好不添加溶剂，以便在后期添加选定的缓冲液，所得分装样本应储存在-20°C下最多几个月，或在-80°C下储存更长时间——长达2年。质量控制方案应表征微生物组成、活力以及代谢功能。
