肠道菌群结构多样性初筛研究

Screening of Intestinal Microflora Diversity

目的:探讨研究肠道菌群结构的初筛方法,为进行有效、高质量的高通量测序奠定基础.方法:用粪便采样器采集健康儿童粪便进行粪便标本预处理后,采用酚—氯仿方法提取样本菌群基因组DNA,并进行16S rDNAV3区片段PCR扩增,扩增产物用变性梯度凝胶电泳法(denaturing gradient gel electrophoresis,DGGE)检测分析.结果:采用该方法成功地扩增出16S rDNA V3区230bp的片段,经DGGE的方法可以比较出不同儿童粪便样本菌群结构的不同.结论:基于16S rDNA V3区的PCR-DGGE技术能够应用于研究肠道微生物多样性,对于肠道微生物的研究起到了初筛的作用.

The screening method of the structure of intestinal flora in children with autism was explored for an effective way of sequencing with high quality and high flux.A sampler was used to collect the fecal samples from healthy children.Total bacteria genomic DNA was extracted from the fecal samples by the method of phenol-chloroform after pre-treatment.For 16S r DNA V3 region PCR amplification,and PCR products were detected and analyzed by 8% denaturing gradient gel electrophoresis（DGGE）.It is found that the method can amplify 16S r DNA V3 specific regions of the 230bp fragment successfully.The detected results by the DGGE method can compare different intestinal bacteria structures between children.It is concluded that the PCR-DGGE technology based on 16S r DNA V3 region can be applied in the study of intestinal microbial diversity,which plays a screening role in intestinal micro-organisms for the study of intestinal microbial.