Comparison of Collection Methods for Fecal Samples in Microbiome Studies

Emily Vogtmann, Jun Chen, Amnon Amir, Jianxin Shi, Christian C. Abnet, Heidi Nelson, Rob Knight, Nicholas Chia, Rashmi Sinha

Research output: Contribution to journalArticlepeer-review

59 Scopus citations


Prospective cohort studies are needed to assess the relationship between the fecal microbiome and human health and disease. To evaluate fecal collection methods, we determined technical reproducibility, stability at ambient temperature, and accuracy of 5 fecal collection methods (no additive, 95% ethanol, RNAlater Stabilization Solution, fecal occult blood test cards, and fecal immunochemical test tubes). Fifty-two healthy volunteers provided fecal samples at the Mayo Clinic in Rochester, Minnesota, in 2014. One set from each sample collection method was frozen immediately, and a second set was incubated at room temperature for 96 hours and then frozen. Intraclass correlation coefficients (ICCs) were calculated for the relative abundance of 3 phyla, 2 alpha diversity metrics, and 4 beta diversity metrics. Technical reproducibility was high, with ICCs for duplicate fecal samples between 0.64 and 1.00. Stability for most methods was generally high, although the ICCs were below 0.60 for 95% ethanol in metrics that were more sensitive to relative abundance. When compared with fecal samples that were frozen immediately, the ICCs were below 0.60 for the metrics that were sensitive to relative abundance; however, the remaining 2 alpha diversity and 3 beta diversitymetrics were all relatively accurate, with ICCs above 0.60. In conclusion, all fecal sample collection methods appear relatively reproducible, stable, and accurate. Future studies could use these collection methods for microbiome analyses.

Original languageEnglish (US)
Pages (from-to)115-123
Number of pages9
JournalAmerican journal of epidemiology
Issue number2
StatePublished - Jan 15 2017


  • Feces
  • Microbiota
  • Specimen collection

ASJC Scopus subject areas

  • Epidemiology


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