一种提取小鼠肠道中活体微生物的方法

An extracting method of alive intestinal microbes from mice

目的建立一种有效的肠道微生物提取方法,用于基于培养方法研究肠道微生物。方法研究NaCl溶液体积、内容物振荡时间、离心转速和高速离心次数对肠道微生物提取数量、质量及其活性的影响。结果在盛有适量玻璃珠的三角瓶中,将肠道内容物与0.85%的NaCl溶液按质量体积比(g∶mL)不大于1∶10进行混合,120 r/min漩涡振荡30 min,500 r/min离心5 min去除沉淀,将上清液分装于EP管中12 000 r/mim高速离心1次,弃去上清,在EP管中加入0.85%NaCl溶液进行漩涡振荡使菌体充分分散,即可得到提取完全、质量高的小鼠肠道微生物。结论该方法能够提取足够多的肠道微生物,并且保持其活性。

Objective To establish an effective method of extraction of intestinal microbs for culture-based researches on intestinal microflora.Methods This experiment studied some factors which could influence the quality,quantity and bioactivity of gut microbes extracted,such as the concentration and volume of NaCl solution,the amount of time intestinal contents were shakened,the speed of centrifuge,and the number of times of high-speed centrifuge.Results The intestinal contents and 0.85% NaCl solution were blended at a weight/volume ratio（g/ml） of no more than 1：10 in a conical flask containing appropriate amount of glass beads,then shaked for 30 min at 120 r/min.After centrifuging at 500 r/min for 5 min,the precipitation was removed and the supernatant was transfered to EP tubes and centrifuged once at 12 000 r/min.Then the supernatant was discarded,and 0.85% NaCl solution was added to the EP tubes.The tubes were vortex-shaked to make the precipitated microbes dispersed thoroughly,resulting in a completely extracted high-quality mice intestinal microbes.Conclusion This method can extract enough amount of intestinal microorganisms with biological activity.