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Use of 16S rRNA Gene-Targeted Group-Specific Primers for Real-Time PCR Analysis of Predominant Bacteria in Human Feces
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16S rRNA gene-targeted group-specific primers were designed and validated for specific detection and quantification of the Clostridium leptum subgroup and the Atopobium cluster. To monitor the predominant bacteria in human feces by real-time PCR, we used these specific primers together with four sets of group-specific primers for the Clostridium coccoides group, the Bacteroides fragilis group, Bifidobacterium , and Prevotella developed in a previous study (T. Matsuki, K. Watanabe, J. Fujimoto, Y. Miyamoto, T. Takada, K. Matsumoto, H. Oyaizu, and R. Tanaka, Appl. Environ. Microbiol. 68: 5445-5451, 2002). Examination of DNA extracted from the feces of 46 healthy adults showed that the C. coccoides group was present in the greatest numbers (log 10 10.3 ± 0.3 cells per g [wet weight] [average ± standard deviation]), followed by the C. leptum subgroup (log 10 9.9 ± 0.7 cells per g [wet weight]), the B. fragilis group (log 10 9.9 ± 0.3 cells per g [wet weight]), Bifidobacterium (log 10 9.4 ± 0.7 cells per g [wet weight]), and the Atopobium cluster (log 10 9.3 ± 0.7 cells per g [wet weight]). These five bacterial groups were detected in all 46 volunteers. Prevotella was found in only 46% of the subjects at a level of log 10 9.7 ± 0.8 cells per g (wet weight). Examination of changes in the population and the composition of the intestinal flora for six healthy adults over an 8-month period revealed that the composition of the flora of each volunteer remained stable throughout the test period.
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“Firstly, the identification of E. coli species was performed by a specific PCR protocol for the 16S rRNA gene using the primers and conditions previously described [51].”
Metadata
- Authors
- Matsuki, T., Watanabe, K., Fujimoto, J., Takada, T., & Tanaka, R.
- Venue
- Applied and Environmental Microbiology
- Publisher
- American Society for Microbiology
- Published
- 2004-12
- Volume
- 70
- Issue
- 12
- Pages
- 7220-7228
- ISSN
- 0099-2240 (ISSN-L)
- 1098-5336
- OpenAlex
- W2014216380
- Cited by
- 655
- References
- 1
- FWCI
- 4.92
- Type
- journal article
- Language
- en
- Open access
- green
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