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目的建立能快速筛查和比较大规模人群肠道中柔嫩梭菌类群组成结构的T-RFLP(末端限制性片段长度多态性)。方法采用T-RFLP技术特异性地分析柔嫩梭菌类群的组成,考察该方法的重复性和灵敏度。用该方法对15个志愿者肠道中的柔嫩梭菌类群进行分析,并与DGGE方法分析的结果进行对比。结果T-RFLP方法重复性高,最低能检测到群落中1%的细菌,其结果与DGGE的分析结果具有较好的一致性。结论本研究建立的柔嫩梭菌类群特异性的T-RFLP方法能够对大量肠道样品中柔嫩梭菌类群的结构进行快速、有效的筛查和比较。
Objective To establish a T-RFLP (Terminal Restriction Fragment Length Polymorphism) that can rapidly screen and compare the compositional structure of the Clostridium difficile in the gut of large-scale population. Methods T-RFLP technique was used to analyze the composition of the Clostridium tenuis and its repeatability and sensitivity. This method was used to analyze the C. difficile population in the gut of 15 volunteers and to compare with the results of DGGE analysis. Results The method of T-RFLP was highly reproducible, and the lowest detection limit was 1% of the bacteria in the community. The results were in good agreement with the results of DGGE analysis. Conclusion The T-RFLP method of C. difficile group established in this study can rapidly and effectively screen and compare the structure of B. flexneri in a large number of intestinal samples.