db/db2型糖尿病模型小鼠肠道多形拟杆菌实时荧光定量PCR标准品制备方法的比较研究  

Comparison of standard sample preparation methods with real time fluorescence quantitative PCR of Bacteroides thetaiotaomicron in intestinal from db/db type 2 diabetic mellitus mice

作　　者：易金阳[1] 王烨[1] 朱明[2] 韩雪[1] 杨珍珍[1] 姬凤彩[3] 郑树涛[4] 毛新民[5] 王丽风[6] 马晓丽[7] 李琳琳[1] 

机构地区：[1]新疆医科大学基础医学院药理学教研室,乌鲁木齐830011 [2]新疆医科大学基础医学院免疫学教研室,乌鲁木齐830011 [3]新疆医科大学实验动物中心,乌鲁木齐830011 [4]新疆医科大学第一附属医院,乌鲁木齐830011 [5]新疆医科大学中医学院,乌鲁木齐830011 [6]新疆医科大学基础医学院生理学教研室,乌鲁木齐830011 [7]新疆医科大学分析测试中心,乌鲁木齐830011

出　　处：《新疆医科大学学报》2012年第6期716-720,724,共6页Journal of Xinjiang Medical University

基　　金：国家自然科学基金(81060068)

摘　　要：目的比较研究db/db 2型糖尿病模型小鼠肠道多形拟杆菌(Bacteroides thetaiotaomicron)实时荧光定量PCR 2种标准品制备方法。方法收集2型糖尿病小鼠db/db模型组和db/m对照组肠道内容物,同时以多形拟杆菌ATCC29148为标准菌株,提取微生物DNA,用16SrRNA V6区多形拟杆菌引物,采用实时荧光定量PCR标准品制备中标准菌株法和正常组高阳性法2种标准品制备方式进行定量检测,比较2种方法的差异性。结果标准菌株法与正常组高阳性法测定db/m对照组和db/db模型组小鼠肠道多形拟杆菌的数量分别为(5.23×106±2.30×106)、(1.52×107±3.23×106)和(5.23×106±2.25×106)、(1.45×107±3.39×106),与db/m对照组相比,db/db模型组多形拟杆菌数量多,差异有统计学意义(P<0.05);标准菌株法与正常组高阳性法对检测多形拟杆菌的差异无统计学意义(P>0.05)。结论实时荧光定量PCR的标准品制备中标准菌株法与正常组高阳性法均可检测出较多的多形拟杆菌,并且2种检测方式都可以对肠道菌群进行检测。Objective The comparative studies for enteric bacteroid content by using real-time fluorescent quantitative PCR with two method of standard preparation in the db/db mice.Methods Collected the intestinal fecal and extract the microbial DNA,and use the V6 District of 16SrRNA bacteroides thetaiotaomicron primers to do the RT-qPCR and assay the content of intestinal Bacteroides thetaiotaomicron.Selected the ATCC29148 bacteroides thetaiotaomicron as a standard strain.Results The number of Bacteroides thetaiotaomicron in type 2 diabetic mellitus mice db/db group and control db/m group by RT-qPCR,respectively,were（5.23×106±2.30×106）,（1.52×107±3.23×106）;（5.23×106±2.25×106）,（1.45×107±3.39×106）.Compared with the control group of db/m,the number of Bacteroides thetaiotaomicron of db/db mice were increased（P〈0.05）.The same results were got from the standard bacteria method and Normal group high positive method.Conclusion Standard bacteria method and Normal group high positive method for quantitative detection of Bacteroides thetaiotaomicron have no significant difference in mice,so the two kinds of detection methods can be used to detect the level of intestinal flora.

关 键 词：db/db鼠 2型糖尿病 多形杆菌 实时荧光定量PCR 

分 类 号：R966[医药卫生—药理学]