Cancer and Inflammation Research, Department of Molecular Medicine, University of Southern Denmark, J. B. Winslows Vej 25.3, DK-5000, Odense C, Denmark.
Institute of Clinical Molecular Biology, Christian Albrechts University of Kiel, Rosalind-Franklin-Str. 12, 24105, Kiel, Germany.
Sci Rep. 2019 Dec 12;9(1):19008. doi: 10.1038/s41598-019-55469-0.
The gut microbiota is recognized as having major impact in health and disease. Sample storage is an important aspect to obtain reliable results. Mostly recommended is immediate freezing, however, this is not always feasible. Faecal occult blood test (FOBT) papers are an appealing solution in such situations, and most studies find these to be applicable, showing no major changes within 7 days storage at room temperature (RT). As fieldwork often requires RT storage for longer periods, evaluation of this is warranted. We performed 16S rRNA gene sequencing of 19 paired faecal samples immediately frozen or kept five weeks and five months at RT on FOBT papers. Alpha-diversity evaluation revealed no effect of FOBT storage, and evaluation of beta-diversity showed that host explained 65% of community variation, while storage method explained 5%. Evaluation of community dispersion and the Firmicutes/Bacteroidetes ratio revealed a larger effect of storage time for fresh-frozen samples. Single taxa evaluation (order-to-genus level) showed significant alterations of four (of 37) genera after five weeks and five genera after five months. When comparing the two timepoints, alterations were only detectable for fresh-frozen samples. Our findings reveal that long term storage on FOBT papers is an applicable approach for microbiota research.
肠道微生物群被认为对健康和疾病有重大影响。样本储存是获得可靠结果的一个重要方面。最推荐的方法是立即冷冻，但这并不总是可行的。在这种情况下，粪便潜血试验（FOBT）试纸是一种有吸引力的解决方案，大多数研究发现这些方法适用，在室温（RT）下储存 7 天内没有明显变化。由于野外工作通常需要在 RT 下储存更长时间，因此有必要对此进行评估。我们对 19 对立即冷冻或在 FOBT 试纸上在 RT 下储存五周和五个月的粪便样本进行了 16S rRNA 基因测序。α多样性评估显示 FOBT 储存没有影响，β多样性评估显示宿主解释了群落变异的 65%，而储存方法解释了 5%。对群落离散度和厚壁菌门/拟杆菌门比值的评估表明，新鲜冷冻样本的储存时间对其影响更大。对 37 个属中的四个属（四个属）进行了单分类群评估（从目到属），五周后有五个属，五个月后有五个属。当比较这两个时间点时，只有新鲜冷冻样本才会检测到变化。我们的研究结果表明，在 FOBT 试纸上进行长期储存是一种适用于微生物组研究的方法。
