探究了不同 DNA 提取方法对人体肠道微生物菌群研究的影响,为今后研究人体肠道微生物时选择提取方法提供参考依据.选择市面上3 种商业化肠道微生物DNA提取试剂盒和1 种实验室改良方法对20份人类粪便样本中微生物DNA进行提取,根据DNA浓度、纯度和微生物多样性比较 4 种DNA提取方法差异.结果表明:Q-试剂盒法(QIAamp PowerFecal Pro DNA Kit,Qiagen)、ET-试剂盒法(ET粪便DNA快速提取试剂盒,IDMBIO)、P-实验室改良法 3 种方法提取的DNA得率较高;方法P获得的OTU数量最多;α多样性分析结果显示 4 种提取方法差异不显著;方法P、方法Q得到肠道微生物组成结构相似度较高,方法ET、方法Y(MolPure Stool DNA Kit,Yeasen)得到肠道微生物组成相似度较高;4种DNA提取方法所得微生物群落构成在门水平和属水平差异明显.每种 DNA 提取方法都有各自提取偏好性菌属,经过综合比较,方法 Q 和方法 ET提取DNA的得率更高、速度更快.
Comparison of DNA Extraction Methods for Human Intestinal Microbial Genomes
The effect of different DNA extraction methods on the study of human intestinal microflora was investigated,in order to provide some reference for DNA extraction links for the community structure research of human gut microorganisms.Three commercial intestinal microbial DNA extraction kits and one improved laboratory method were selected to extract microbial DNA from 20 human feces samples,and the differences of four DNA extraction methods were compared according to DNA concentration,purity and microbial diversity.The results showed that the Q-commercial kit(QIAamp PowerFecal Pro DNA Kit,Qiagen),ET-commercial kit(ET fecal DNA rapid extraction kit,IDMBIO)and P-the improved method had higher DNA yield;the method of P had the largest OTU numbers;there was no significant difference among the four extraction methods in α diversity analysis;methods of P and Q obtained higher similarity of species composition structure,and methods of ET and Y(MolPure Stool DNA kit,Yeasen)obtained higher recognition of species composition;the composition of microbial community obtained by four DNA extraction methods was obviously different at the level of phylum and genus.Each DNA extraction method had its own bacterial bias.The methods of Q and ET had higher extraction rate and shorter operation time in a comprehensive comparison of microorganism DNA extraction.
DNA extractionNext generation sequencingAmplification sequencingMicroflora
万方数据
维普
