Wang M, Ahrné S, Antonsson M, Molin G
Laboratory of Food Hygiene, Department of Food Technology, Engineering and Nutrition, P.O. Box 124, Lund SE-22100, Sweden.
J Microbiol Methods. 2004 Oct;59(1):53-69. doi: 10.1016/j.mimet.2004.06.002.
The fecal microbiota of two healthy Swedish infants was monitored over time by terminal restriction fragment length polymorphism (T-RFLP) analysis of amplified 16S rRNA genes. Principal component analysis (PCA) of the T-RFLP profiles revealed that the fecal flora in both infants was quite stable during breast-feeding and a major change occurred after weaning. The two infants had different sets of microbiota at all sampling time points. 16S rDNA clone libraries were constructed and the predominant terminal restriction fragments (T-RFs) were identified by comparing T-RFLP patterns in the fecal community with that of corresponding 16S rDNA clones. Sequence analysis indicated that the infants were initially colonized mostly by members of Enterobacteriaceae, Veillonella, Enterococcus, Streptococcus, Staphylococcus and Bacteroides. The members of Enterobacteriaceae and Bacteroides were predominant during breast-feeding in both infants. However, Enterobacteriaceae decreased while members of clostridia increased after weaning. T-RFLP in combination with PCA and 16S rRNA gene sequencing was shown to be an effective strategy for comparing fecal microbiota in infants and pointing out the major changes.
通过对扩增的16S rRNA基因进行末端限制性片段长度多态性（T-RFLP）分析，对两名健康瑞典婴儿的粪便微生物群进行了长期监测。T-RFLP图谱的主成分分析（PCA）显示，两名婴儿的粪便菌群在母乳喂养期间相当稳定，断奶后发生了重大变化。在所有采样时间点，两名婴儿的微生物群各不相同。构建了16S rDNA克隆文库，并通过比较粪便群落中的T-RFLP模式与相应16S rDNA克隆的模式来鉴定主要的末端限制性片段（T-RFs）。序列分析表明，婴儿最初主要由肠杆菌科、韦荣氏菌属、肠球菌属、链球菌属、葡萄球菌属和拟杆菌属的成员定殖。两名婴儿在母乳喂养期间，肠杆菌科和拟杆菌属的成员占主导地位。然而，断奶后肠杆菌科减少，而梭菌属的成员增加。T-RFLP与PCA和16S rRNA基因测序相结合被证明是比较婴儿粪便微生物群并指出主要变化的有效策略。
