The reproducibility and stability of stool microbiota during collection and transport of stool specimen
Year: 2020
Type: conference-abstract
Abstract: Background/Aim: The role of gut microbiome in epidemiologic research is increasingly recognized and with it the number of studies analyzing stool specimen is rapidly expanding. However, currently there are no standardized protocols for collecting, transporting and storing stool specimen in population-based environmental research covering a large geographic area. For home data collection, it takes between 12-100 hours from stool collection to time of sample processing in the lab. The aim of the study is to test the stability and reproducibility of stool microbiota during the transit period and provide guidance on collecting stool in population-based studies. Methods: Fresh stool was collected from 12 volunteers. Samples were aliquoted into cryovials within 30 min of collection. Three aliquots from each sample were frozen immediately (time 0) and remaining aliquots were kept at 4oC for 6, 24, 48, 72 and 96 hours followed by storage at -80oC. Three aliquots per sample were analyzed for each time point to assess reproducibility. The composition of stool microbiota was assessed by DNA sequencing of 16s rRNA V4 region. Results: The intraclass correlation coefficient (ICC) for repeated measures was between 0.6-1.0 for α-diversity measures ACE, Chao1, Shannon, and Inverse Simpson. The ICC for relative abundance of four major phyla (Bacteroidetes, Firmicutes, Proteobacteria and Actinobacteria) between time 0 and 6 hours at 4oC was 0.9-1.0. The correlation remained excellent (0.9-1.0) for Proteobacteria and Actinobacteria, but was moderate (ICC from 0.6 to 0.7) for Bacteroidetes and Firmicutes between 24-96 hours at 4oC. Conclusions: Results suggest that a) the moderate correlation of repeated assessments from some of the samples suggests that stool specimen should be homogenized before taking aliquots, and b) stool microbiota is relatively stable at 4oC for up to 96 hours. However, some bacterial taxa had altered relative abundance suggesting they are more sensitive to storage conditions.
Source: ISEE Conference Abstracts
Institution: University of Wisconsin–Madison
Language: English
Topic: Gut microbiota and health
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: diamond
OpenAlex ID: W3170790693