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

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(P0.05).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.