Comparison of Bacterial Communities in the Throat Swabs from Healthy Subjects and Pharyngitis Patients by Terminal Restriction Fragment Length Polymorphism

Terminal restriction fragment length polymorphism (T-RFLP) analysis was applied to characterize bacterial flora present in the throats of healthy subjects and pharyngitis patients. The 16S rRNA genes of bacteria present in throat metagenome were amplified by PCR with 6-carboxy-fluorescein (6-FAM)-la...

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Veröffentlicht in:	Applied biochemistry and biotechnology Jg. 167; H. 5; S. 1459 - 1473
Hauptverfasser:	Balaji, Kannan, Thenmozhi, Ramalingam, Sundaravadivel, Marimuthu, Pandian, Shunmugiah Karutha
Format:	Journal Article
Sprache:	Englisch
Veröffentlicht:	New York Springer-Verlag 01.07.2012 Springer Nature B.V
Schlagworte:	Bacteria Bacteria - classification Bacteria - genetics Bacteria - isolation & purification Bacteria - pathogenicity bacterial communities Bacteriology Biochemistry Biodiversity Biotechnology Chemistry Chemistry and Materials Science classification Community structure computer software Enzymes Flora genes genetics Genomes Health Humans isolation & purification Metagenome Metagenome - genetics metagenomics microbiology pathogenicity Pathology patients Pharyngitis Pharyngitis - microbiology Pharynx Pharynx - microbiology Phylogeny Polymerase chain reaction Polymorphism Polymorphism, Restriction Fragment Length restriction fragment length polymorphism ribosomal DNA ribosomal RNA RNA, Bacterial RNA, Bacterial - genetics RNA, Ribosomal, 16S RNA, Ribosomal, 16S - genetics Species diversity Species richness Streptococcus pyogenes throat uncertainty Normal flora Restriction enzymes Pharyngitis Metagenome T-RFLP
ISSN:	0273-2289, 1559-0291, 1559-0291
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Zusammenfassung:	Terminal restriction fragment length polymorphism (T-RFLP) analysis was applied to characterize bacterial flora present in the throats of healthy subjects and pharyngitis patients. The 16S rRNA genes of bacteria present in throat metagenome were amplified by PCR with 6-carboxy-fluorescein (6-FAM)-labeled universal forward primer (27 F) and a universal reverse primer (1513R). The 16S rDNAs were digested with restriction enzymes with 4-bp recognition sites ( Msp I or Rsa I) and analyzed by using an automated DNA sequencer. T-RFLP patterns were numerically analyzed using computer programs. From analysis of the throat bacterial community, patterns derived from Msp I and Rsa I digested samples of healthy subjects and pharyngitis patients were grouped into different clusters, though Rsa I digested samples showed some uncertainty. Pharyngitis throats generated an average species richness of 9 [±2.1 (SD)] and 10 (±2.9) for Msp I and Rsa I digests, respectively, whereas healthy throats generated 6.3 (±1.2) and 6.1 (±1.5) in Msp I and Rsa I digests, respectively. These results suggest that samples from pharyngitis patients contain an unexpected diversity of causative bacteria. The pharyngitis throats were colonized with a rich diversity of bacterial species than that of healthy throats. Using T-RFLP, we are able to detect a model bacterium, Streptococcus pyogenes SF370, and T-RF patterns were consistent with the Streptococcal T-RFLP patterns. Our study indicates that T-RFLP analysis is useful for the assessment of diversity of throat bacterial flora and rapid comparison of the community structure between subjects with and without pharyngitis.
Bibliographie:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 14 content type line 23 ObjectType-Article-1 ObjectType-Feature-2
ISSN:	0273-2289 1559-0291 1559-0291
DOI:	10.1007/s12010-011-9508-4