PCR-DGGE法在慢性胃炎病人舌苔菌群结构研究中的应用

Application of PCR-DGGE in the Analysis of Structure of Bacteria Colony from Tongue Coating in Gastritis Patients

在早间未刷牙和进食的情况下,刮取胃炎病人与正常人舌苔,去除杂质,分离菌体,采用酚/氯仿法抽提细菌基因组DNA,并对其中16S rDNA V3可变区进行聚合酶链式反应(PCR)扩增和变性梯度凝胶电泳(DGGE)测定.用Bionumerics软件对DGGE分子指纹图谱进行舌苔菌群结构相似性分析.实验结果表明,采用该方法成功地扩增出16S rDNA V3区片段,为230 bp.DGGE分子指纹图谱结果表明,正常人的舌苔菌群最高相似性为0.74,胃炎病人的舌苔菌群的最高相似性为0.52,正常人的舌苔菌群与胃炎病人的舌苔菌群相似性最高为0.38,即胃炎病人舌苔菌群结构发生了变化.

Tongue coating of gastritis patient and normal persons were scraped without cleaning teeth and no food in the morning.Impurities were eliminated and the bacteria was separated.The total microbial genome DNA of each sample was extracted by phenol-chloroform,V3 regions of 16S rDNA was amplified with PCR,and PCR products were analyzed by DGGE.The similarity of bacteria structure was analyzed with Bionumerics.The result showed that V3 regions of 16S rDNA was extracted,230 bp.DGGE suggested that the highest similarity was 0.74 in normal persons,that of gastritis patients was 0.52,and that of normal persons and gastritis patients was 0.38.Therefore,some variation existed in structure of bacteria colony in tongue coating of gastritis patient.