Abstract
BackgroundThe number of large-scale studies on the gut microbiota in human cohorts is rapidly increasing. However, the few and expensive options for storage of fecal samples at room temperature have been an obstacle for large-scale metagenomic studies and the development of clinical/commercial personal metagenomic sequencing.ResultsIn this study, we systematically tested a novel N-octylpyridinium bromide-based fecal sample preservation method and compared it with other currently used storage methods. We found that the N-octylpyridinium bromide-based method enabled preservation of the bacterial composition in fecal samples transported and stored at room temperature for up to at least 14 days.ConclusionsWe describe a novel chemical stabilizer that allows cost-effective transportation and storage at room temperature for several days with preservation of bacterial composition. This method will facilitate sample collection even in remote area and also enable transport via normal commercial transportation routes.
Highlights
The number of large-scale studies on the gut microbiota in human cohorts is rapidly increasing
Distance and dissimilarity between samples stored in the Noctylpyridinium bromide (NOPB)-based and freshly extracted samples were significantly lower than those observed in analyzing the non-stabilized samples, the correlation coefficient measured by the Spearman index did not show significant difference between the two protocols (Additional file 1: Figure S1)
Compared with the corresponding fresh samples, aliquots stored in the NOPB-based stabilizer reagent and the samples kept at − 80 °C and transported in dry ice showed significantly lower dissimilarity in relation to relative gene abundances and microbiome composition than those in the non-stabilized samples after 7-day storage (Fig. 2b, c)
Summary
The number of large-scale studies on the gut microbiota in human cohorts is rapidly increasing. The few and expensive options for storage of fecal samples at room temperature have been an obstacle for large-scale metagenomic studies and the development of clinical/commercial personal metagenomic sequencing. Fecal samples are widely used in metagenomic studies and are generally required to be immediately stored at − 20 °C or below. The aliquots were stored or transported at room temperature using different schemes before extraction. After storage and/or transportation, all aliquots were sequenced, and each was compared to the corresponding freshly extracted sample. Extracted DNA from all samples was sequenced on Illumina HiSeq 4000, and a subset was sequenced on the BGISEQ-500 instrument
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