T-RFLP技术在人肠道柔嫩梭菌类群的组成分析中的应用

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。

The application of T-RFLP in analyzing Clostridium leptum group in human gut microbiota

Objective To establish a group-specific Terminal Restriction Fragment Length Polymorphisms（T-RFLP） method to analyze the structure of Clostridium leptum group in human gut samples collected from large-scale populations.Method The reproducibility and sensitivity of Clostridium leptum group specific T-RFLP was determined.T-RFLP and DGGE were used to analyze the fecal Clostridium leptum subgroup of 15 human subjects and the results were compared to evaluate the consistency of the two techniques.Result The Clostridium leptum group-specific T-RFLP method was shown to be of high reproducibility and had the detection limit down to 1%.The result of T-RFLP was in good coherence with that of DGGE.Conclusion This group-specific T-RFLP method could be used to efficiently screen and compare the structure of Clostridium leptum group in large quantity of fecal samples.