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T-RFLP combined with principal component analysis and 16S rRNA gene sequencing: an effective strategy for comparison of fecal microbiota in infants of different ages

Wang, Mei LU ; Ahrné, Siv LU ; Antonsson, M and Molin, Göran LU (2004) In Journal of Microbiological Methods 59(1). p.53-69
Abstract
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... (More)
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. (C) 2004 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
16S rDNA clone library, T-RFLP, Infant fecal microbiota, PCA
in
Journal of Microbiological Methods
volume
59
issue
1
pages
53 - 69
publisher
Elsevier
external identifiers
  • wos:000223935300007
  • scopus:4344626157
ISSN
1872-8359
DOI
10.1016/j.mimet.2004.06.002
language
English
LU publication?
yes
id
b8316a50-0dbb-4fcd-9b27-b6e22adef1e1 (old id 139297)
date added to LUP
2007-07-19 11:30:04
date last changed
2017-11-12 03:18:30
@article{b8316a50-0dbb-4fcd-9b27-b6e22adef1e1,
  abstract     = {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. (C) 2004 Elsevier B.V. All rights reserved.},
  author       = {Wang, Mei and Ahrné, Siv and Antonsson, M and Molin, Göran},
  issn         = {1872-8359},
  keyword      = {16S rDNA clone library,T-RFLP,Infant fecal microbiota,PCA},
  language     = {eng},
  number       = {1},
  pages        = {53--69},
  publisher    = {Elsevier},
  series       = {Journal of Microbiological Methods},
  title        = {T-RFLP combined with principal component analysis and 16S rRNA gene sequencing: an effective strategy for comparison of fecal microbiota in infants of different ages},
  url          = {http://dx.doi.org/10.1016/j.mimet.2004.06.002},
  volume       = {59},
  year         = {2004},
}