한국인의 정상 치주조직 및 치주질환 병소의 치면세균막에서 Fusobacterium nucleatum의 발현빈도 비교

Abstract

Fusobacterium nucleatum is a gram-negative anaerobic rod implicated in the pathogenesis of periodontal diseases. F. nucleatum has five subspecies and the genetic- and phenotypic-characteristics are variable among the subspecies. These observations suggest that the detection frequency of subspecies of F. nucleatum might be different according to the degree of periodontal diseases. The purpose of this study was to investigate the distribution of subspecies of F. nucleatum in the subgingival plaques from the periodontal healthy region and periodontitis (gingivitis, periodontitis, and periimplantitis) lesion. In addition, we evaluate the specificity of Fu4-F38 and Fu4-R1189 primers, introduced in Shin (PhD paper in Graduate School of Chosun University, 2005), whether they can be useful in the detection and identification of F. nucleatum. The subgingival plaques from total 346 tooth sites (113 persons) were collected; 88 tooth sites (44 persons) of healthy region, 193 sites (36 persons) of gingivitis lesion, 45 sites (22 persons) of periodontitis lesion, and 22 sites (11 persons) of periimplantitis subjects. The identification of the isolates, growing on the F. nucleatum-selective media, at the species level by 16S rDNA sequencing comparison method. For the identification of the F. nucleatum at the subspecies level, Fu4 DNA probe-like region were amplified from the genomic DNAs of clinical isolated from the subgnigival plaques and sequences of them. The species-specificity of the Fu4-F38 and Fu4-R1189 primers to the F. nucleatum were evaluated by PCR with the genomic DNAs from 7 other Fusobacterium species. The 56 strains of F. nucleatum and 7 strains of F. periodonticum were isolated from the subgingival plaques. The five subspecies of F. nucleatum exhibited relatively high levels of sequence similarity(the hemology range is 97.8 to 99.7%) by 16S rDNA sequences. On the other hand, the Fu4 DNA probe-like sequences were significantly different among the subspecies of F. nucleatum (the hemology range is 89.7 to 99.0%). F. nucleatum subsp. vincentii/fusiforme was associated with healthy region and both F. nucleatum subsp. nucleatum and F. nucleatum subsp. animalis were associated with periodontitis lesion. The PCR data showed that Fu4-F38 and Fu4-R1189 primers were specific to the F. nucleatum. These results suggest that distribution of 5 subspecies of F. nucleatum were different in the according to the degree of periodontal diseases. In the further, study, new methodology must be developed to the detection of F. nucleatum at the subspecies level and try to reinvestigate the detection frequency of subspecies of F. nucleatum with the more samples.