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目的采用常规菌群分析方法和ERIC-PCR技术对正常小鼠和抗生素相关性腹泻小鼠模型分别进行肠道菌群检测,结合细菌培养和DNA指纹图谱检测结果分析小鼠肠道内主导菌群数量和种类的改变情况,建立利用ERIC-PCR技术分析小鼠肠道菌群失调的检测方法。方法先利用常规菌群分析方法鉴定正常小鼠和抗生素相关性腹泻小鼠的菌群状况,再提取其基因组DNA,最后以肠杆菌科基因间重复序列(ERIC)为模板,利用ERIC-PCR方法获得正常小鼠和模型小鼠的肠道菌群指纹图谱,与常规菌群分析结果作比较,验证ERIC-PCR技术的准确性。结果常规菌群分析结果表明四种优势菌群在数量上出现明显的变化,证实造模成功。经ERIC-PCR技术成功获得两组小鼠粪便基因组DNA图谱,两组间呈现具有一定对比性的特异性指纹图谱。结论从小鼠粪便基因组经ERIC-PCR后的图谱中特异性条带的分布、数目和亮度来看,能说明肠道菌群的分布状况存在明显差异,结合常规菌群分析方法作对比,说明ERIC-PCR技术是一种分析小鼠肠道菌群失调高效快捷的检测方法。
Objective To detect the intestinal microflora in normal mouse and antibiotic-associated diarrhea model by routine bacterial population analysis and ERIC-PCR, and to analyze the number of dominant gut flora in mouse intestine by bacterial culture and DNA fingerprinting And types of changes in the establishment of the use of ERIC-PCR analysis of intestinal disorders in mice detection method. Methods Firstly, the flora of normal mice and antibiotic-associated diarrhea mice were identified by routine bacterial population analysis, and then the genomic DNA was extracted. Finally, the sequence of ERIC was used as the template and ERIC-PCR method The gut microflora fingerprints of normal and model mice were obtained and compared with the results of routine bacterial population analysis to verify the accuracy of ERIC-PCR. Results The results of routine bacterial population analysis showed that there were obvious changes in the number of the four dominant bacteria, confirming the success of the model. The genomic DNA of feces from two groups of mice was successfully obtained by ERIC-PCR. The two groups showed a certain specific fingerprint. Conclusion The distribution, number and brightness of specific bands in the stool genome of mice after ERIC-PCR showed that the distribution of intestinal flora is significantly different. Combined with conventional bacterial population analysis, ERIC -PCR technology is a rapid and efficient assay for the analysis of intestinal flora in mice.