CAJ | 학술논문

目的：利用重复共有序列（ERIC）和变性梯度凝胶电泳（DGGE）两种分子指纹图谱技术对2型糖尿病患者肠道菌群结构特征分析，探讨2型糖尿病与肠道菌群的相关性及对两种方法的评价。方法收集8例健康人和7例2型糖尿病患者粪便样本，提取粪便菌群总 DNA，采用 ERIC-PCR 和 DGGE-PCR 分子指纹图谱技术对两组人群肠道菌群结构分析，比较其多样性、相似性等生态学特征。结果与健康对照组比较，2型糖尿病患者肠道菌群图谱条带和 Shannon-Wiener 多样性指数降低，但无统计学意义；组内相似性降低差异有统计学意义（P ＜0.05），菌群结构发生变化；两种指纹图谱技术均能直观反映肠道菌群结构特征，ERIC 方法简便，反映菌群多样性良好，但是实验影响因素较多，不可切胶测序；DGGE 能较好反映菌群多样性、相似性等生态学特征，而且可选择条带切胶测序。结论2型糖尿病患者肠道菌群组成结构发生改变，糖尿病的发生与肠道菌群有一定相关性；ERIC 和 DGGE 是研究肠道菌群分辨效率高、重复性好的指纹图谱技术， DGGE 并可进行切胶测序比对鉴定细菌，二者可结合使用。
목적：이용중복공유서렬（ERIC）화변성제도응효전영（DGGE）량충분자지문도보기술대2형당뇨병환자장도균군결구특정분석，탐토2형당뇨병여장도균군적상관성급대량충방법적평개。방법수집8례건강인화7례2형당뇨병환자분편양본，제취분편균군총 DNA，채용 ERIC-PCR 화 DGGE-PCR 분자지문도보기술대량조인군장도균군결구분석，비교기다양성、상사성등생태학특정。결과여건강대조조비교，2형당뇨병환자장도균군도보조대화 Shannon-Wiener 다양성지수강저，단무통계학의의；조내상사성강저차이유통계학의의（P ＜0.05），균군결구발생변화；량충지문도보기술균능직관반영장도균군결구특정，ERIC 방법간편，반영균군다양성량호，단시실험영향인소교다，불가절효측서；DGGE 능교호반영균군다양성、상사성등생태학특정，이차가선택조대절효측서。결론2형당뇨병환자장도균군조성결구발생개변，당뇨병적발생여장도균군유일정상관성；ERIC 화 DGGE 시연구장도균군분변효솔고、중복성호적지문도보기술， DGGE 병가진행절효측서비대감정세균，이자가결합사용。
Objective To explore the characteristics of intestinal Microbiota in T2DM patients by two molecular fingerprint technologies,and investigate the correlation of intestinal microbiota and T2DM,and evaluate the application value of two fin-gerprint technologies.Methods Fecal samples of 8 healthy groups and 7 diabetes patients were collected.Then the total DNA of gut microbiota was extracted.Through the analysis of products by two molecular fingerprints of ERIC-PCR and DGGE-PCR,ecological characteristics of diversity and similarity of gut microbiota were obtained in healthy groups and dia-betes patients.Results Compared to healthy groups,the number of bands and Shannon-Wiener index of gut microbiota in di-abetes patients was decreased but no statistical significance.The similarity in patients group was declining(P <0.05),and the construction of gut microbiota was inclined to differ.Two fingerprint technologies of ERIC and DGGE could directly re-flect the diversity of gut microbiota and were the modern molecular biological techniques without depending on cultivation. ERIC was simple and convenient,had a better reflection of microbial diversity,but gel band cutting and regarded asa proper approach with higher diffraction efficiency and excellent repetition to studysequencing couldn’t be performed since there were more influencing factors on the experiment.DGGE could better reflect the ecological characteristics such as microbial diversity and similarity,and selecting bands,gel band cutting and sequencing could be done.Conclusion The composition and construction of gut microbiota in diabetes patients were changed,which suggests the occurrence of the disease had the correlation with gut microbiota.ERIC and DGGE is regarded as a proper approach with higher diffraction efficiency and ex-cellent repetition to study intestinal microbiota,but also gel band cutting,sequencing,bacteria identification can be performed by DGGE,both can be used in combination.