Fecal Microbiota Transplantation and Detection of Prevalence of IgA-Coated Bacteria in the Gut

J Vis Exp. 2020 Jul 23:(161). doi: 10.3791/60772.

Abstract

Gut microbiota exert pleiotropic roles in human health and disease. Fecal microbiota transplantation (FMT) is an effective method to investigate the biological function of intestinal bacteria as a whole or at the species level. Several different FMT methods have been published. Here, we present an FMT protocol that successfully depletes gut microbiota in a matter of days, followed by transplantation of fecal microbiota from fresh or frozen donor intestinal contents to conventional mice. Real time-PCR is applied to test the efficacy of bacterial depletion. Sequencing of the 16S ribosomal RNA (rRNA) is then applied to test the relative abundance and identity of gut microbiota in recipient mice. We also present a flow cytometry-based detection method of immunoglobulin A (IgA)-coated bacteria in the gut.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Fecal Microbiota Transplantation / methods*
  • Gastrointestinal Microbiome / immunology*
  • Humans
  • Immunoglobulin A / metabolism*
  • Mice
  • Prevalence

Substances

  • Immunoglobulin A