摘要
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 establish a method based on ERIC-PCR technique for detection of mouse dysbacteriosis. Methods The intestinal flora of normal mice and mouse models of antibiotic-associated diarrhea were detected by using conventional culture methods, then the genomic DNA was extracted. With enterobacteria repetitive intergenic consensus sequence （ERIC） as the template, the fingerprint spectra of intestinal flora of normal mice and mouse models were obtained with ERIC-PCR technology. The results were compared with those of conventional methods to verify the accuracy of ERIC PCR method. Results Conventional detection showed that the eoums of four dominant bacteria had a significant change, suggesting that the model group was established successfully. The genomic DNA patterns obtained from mouse droppings by using ERIC PCR technique presented specific fingerprints which were comparative between the two groups. Conclusion The spe cific fingerprint bands revealed that there was significant difference in the distribution of intestinal microflora between the two groups. In combination with conventional method, the ERIC-PCR technology is a fast and efficient analytical method for detecting mouse intestinal flora.
出处
《中国微生态学杂志》
CAS
CSCD
2016年第12期1386-1388,1392,共4页
Chinese Journal of Microecology
