Kazuhiko Nakano, Atsuo Amano and Takashi Ooshima
48
(formerly
Actinobacillus
)
actinomycetemcomitans
(Aa),
Capnocytophaga ochracea
(Co),
Capnocytophaga sputigena
(Cs) and
Treponema denticola
(Td) [47-49], based on previous
reports showing that their distributions in periodontitis patients were significantly different
from those in periodontally healthy subjects [50-55]. Table 4 lists the primer sets specific for
each species used in our studies, all of which were checked for specificity as well as
sensitivityand was reported to range from 10-100 cells in the original studies [39-43]. Figure
18 shows an example of the results for the detection of 10 periodontal bacterial species by
PCR with species-specific sets of primers. There were two species identified in the saliva
specimen of subject A, whereas 6 species were detected in the specimen taken from subject
B.
Figure 18. Detection of periodontal bacterial species in oral specimens by PCR methods. The left
pictures show the two representative cases (subjects A and B) and the right images are stained
electrophoresis gels for identification of the species in the specimens taken from each case. ―M‖ and
―PC‖ indicate molecular marker and positive control, respectively, and arrows
indicate the positive
reactions.
2) Distribution of Periodontal Bacteria in Children
There have been few reports describing the results of the distribution of periodontal
bacterial species in children and adolescents when we initiated this study. It was generally
known that children with periodontitis are rarely encountered, which might be one of the
reasons for the few studies analyzing their distribution. Therefore, we decided to analyze the
distribution of the 10 selected bacterial species in the dental plaqueand saliva specimens in
children and adolescents who came to our clinic. Dental plaque specimens were collected
from the buccal-mesial sulcus of the first molar or second primary molar in the right upper
quadrant of 119 systemically healthy children (56 boys and 63 girls) aged 2 to 13 years old,
who showed negligible periodontal inflammation, and their whole expectorated saliva
specimens were also collected [47, 48]. The total numbers of dental plaque and saliva
Kazuhiko Nakano, Atsuo Amano and Takashi Ooshima
50
detection using 10 species-specific primer sets showed that approximately 15% of the
subjects were negative for any of those 10 species. The total numbers of the detected species
increased gradually with age until 5 years old and then reached a plateau after the mixed
dentition period. The detection rates for most of the species in saliva specimens were
significantly higher than those in dental plaque specimens.
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