Zhang Qiuping, Lu Qian, Luo Yubin
Department of Rheumatology and Immunology, West China Hospital, Sichuan University.
Department of Biomedical Engineering, College of Engineering and Applied Sciences, Nanjing University.
J Vis Exp. 2020 Jul 23(161). doi: 10.3791/60772.
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.
肠道微生物群在人类健康和疾病中发挥着多效性作用。粪便微生物群移植（FMT）是一种研究肠道细菌整体或物种水平生物学功能的有效方法。已经发表了几种不同的FMT方法。在此，我们提出一种FMT方案，该方案能在数天内成功清除肠道微生物群，随后将新鲜或冷冻供体肠道内容物中的粪便微生物群移植到常规小鼠体内。应用实时聚合酶链反应（Real time-PCR）检测细菌清除的效果。然后应用16S核糖体RNA（rRNA）测序检测受体小鼠肠道微生物群的相对丰度和种类。我们还提出了一种基于流式细胞术的肠道中免疫球蛋白A（IgA）包被细菌的检测方法。
