Application of terminal RFLP analysis to characterize oral bacterial flora in saliva of healthy subjects and patients with periodontitis

• DOI: 10.1099/jmm.0.04991-0
• Publication Date: 2002-12-18T19:14:57Z
• Authors: Mitsuo Sakamoto; Yasuo Takeuchi; Makoto Umeda; Isao Ishikawa; Yoshimi Benno
• Keywords: Oral microbiology; Bacterial taxonomy

Terminal restriction fragment-length polymorphism (T-RFLP) analysis was applied to characterize oral bacterial flora in saliva from 18 healthy subjects and patients with periodontitis. The 16S rRNA genes (rDNAs) of bacteria spirochaetes were amplified by PCR a 6′carboxy-fluorescein (6-FAM)-labelled universal forward primer (27F) reverse (1492R) or the Spirochaeta-selective primer. rDNAs digested enzymes 4 bp recognition sites (HhaI MspI) analysed using an automated DNA sequencer. T-RFLP patterns numerically computer program. From community, derived periodontally periodontitis grouped into different clusters, though some uncertainty. Samples tended cluster their respective types (aggressive chronic periodontitis), although this not very clear. Analysis spirochaetal community showed that more as compared community. These results suggest samples contain unexpected diversity. two over 5-week period host-specific relatively stable flora. Our study indicates is useful for assessment diversity rapid comparison structure between without