Sample storage conditions significantly influence faecal microbiome profiles.
Sample storage conditions significantly influence faecal microbiome profiles.
复制标题
DOI：
10.1038/srep16350
复制
发表时间：
2015-11-17
影响因子：
4.6
通讯作者：
Rogers GB
中科院分区：
文献类型：
--
作者：
Choo JM;Leong LE;Rogers GB
基于测序的人类粪便微生物群研究越来越普遍。样品材料的适当储存对于避免微生物群落组成中引入采集后偏差至关重要。快速冷冻至-80°C 通常被认为是最佳实践。然而，这在许多研究中是不可行的，特别是那些涉及在参与者家中采集样本的研究。我们确定了一系列稳定和储存策略相对于冷冻至-80°C 维持粪便微生物群落结构组成的程度。相对于冷冻，对 4°C 冷藏、环境温度储存以及几种常见防腐缓冲液（RNAlater、OMNIgene.GUT、Tris-EDTA）的使用进行了评估。储存 72 小时后，通过 16S rRNA 扩增子测序评估粪便微生物组成。冷藏与粪便微生物群多样性或组成没有显着改变相关。然而，与-80°C 对照样品相比，使用其他条件储存的样品显示出很大的差异。除了冷藏之外，使用 OMNIgene.GUT 导致的变化最小，而最大的变化出现在 Tris-EDTA 缓冲液中储存的样品中。市售的 OMNIgene.GUT 试剂盒可以在无法冷藏和冷链运输的情况下提供重要的替代方案。
Sequencing-based studies of the human faecal microbiota are increasingly common. Appropriate storage of sample material is essential to avoid the introduction of post-collection bias in microbial community composition. Rapid freezing to −80 °C is commonly considered to be best-practice. However, this is not feasible in many studies, particularly those involving sample collection in participants’ homes. We determined the extent to which a range of stabilisation and storage strategies maintained the composition of faecal microbial community structure relative to freezing to −80 °C. Refrigeration at 4 °C, storage at ambient temperature, and the use of several common preservative buffers (RNAlater, OMNIgene.GUT, Tris-EDTA) were assessed relative to freezing. Following 72 hours of storage, faecal microbial composition was assessed by 16 S rRNA amplicon sequencing. Refrigeration was associated with no significant alteration in faecal microbiota diversity or composition. However, samples stored using other conditions showed substantial divergence compared to −80 °C control samples. Aside from refrigeration, the use of OMNIgene.GUT resulted in the least alteration, while the greatest change was seen in samples stored in Tris-EDTA buffer. The commercially available OMNIgene.GUT kit may provide an important alternative where refrigeration and cold chain transportation is not available.