Molecular analysis of bacteria in periodontitis: evaluation of clone libraries, novel phylotypes and putative pathogens

Abstract

Subgingival plaque samples were obtained from 26 subjects with advanced periodontal lesions. Bacterial diversity was analysed by amplification of the 16S rRNA genes with two different primer sets, and subsequent cloning and sequencing. A total of 578 sequences was analysed after the exclusion of chimeras. The authors found 148 phylotypes with the clone library 27f/519r (number of clones n5322; coverage, C566 %) and 75 phylotypes with the clone library 515f/1525r (n5256; C584 %). Comparative sequence analysis revealed that 70 % of all of the analysed sequences showed a similarity of at least 99 % to sequences deposited in public databases. The classes Actinobacteria, Bacilli, Bacteroidetes, Clostridia, Deferribacteres, Flavobacteria, Fusobacteria, Mollicutes, Spirochaetes and all classes of the Proteobacteria were represented. Sequences that were at least 99 % identical to Porphyromonas gingivalis, Filifactor alocis and Treponema socranskii were present in at least one-third of the patients. Libraries generated with the two PCR primer pairs differed significantly in their representation of the families Porphyromonadaceae, Prevotellaceae, Fusobacteriaceae, Eubacteriaceae, Streptococcaceae and Acidaminococcaceae. A total of 14 sequences exhibited less than 97 % identity to sequences published previously and were assigned to six different families within the phyla Bacteroidetes and Firmicutes. Twelve of 20 putative pathogens were recovered, which were recently proposed to be associated with periodontitis.

Application of substrate utilization patterns and terminal restriction fragment length polymorphism analysis to characterize the oral bacterial community of healthy subjects and patients with periodontitis

“…The present results indicated that the TRFLP patterns derived from patients with periodontitis were significantly different from the patterns of periodontally healthy subjects. The data confirm findings from previous studies, which reported that the saliva from patients with periodontitis contains a special bacterial diversity (9,19). Notably, the present TRFLP results also suggested that the age of the subjects had a significant effect on the oral bacterial flora.…”

Jonquetella anthropi gen. nov., sp. nov., the first member of the candidate phylum ‘Synergistetes’ isolated from man

“…To date, bacteria belonging to the phylum 'Synergistetes' have been found in a large range of ecosystems, including the guts of mammals (Godon et al, 2005). Culture-independent studies have demonstrated their presence in man during endodontic or periodontic infections, in dental caries (Munson et al, 2002(Munson et al, , 2004Siqueira & Rôças, 2005, 2007Hutter et al, 2003), in subgingival plaque (Paster et al, 2001), in faeces (Horz et al, 2006) and in vaginal flora (A. Damay, H. Marchandin, L. Roudière, C. Teyssier, H. Dechaud, H. Jean-Pierre, E. Jumas-Bilak, unpublished results). However, only eight human isolates have been reported to date (Munson et al, 2002;Warren et al, 2005;Horz et al, 2006).…”

Cytokine production by leukocytes in patients with periodontitis

“…The gingival sulcus is home to a complex microbial ecosystem [17]. However, the presence of periodontopathogens is required but not sufficient for disease initiation [18].…”