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Clonal Analysis of the Microbiota of Severe Early Childhood CariesKanasi E.a, c · Dewhirst F.E.a, d · Chalmers N.I.a · Kent, Jr. R.b, c · Moore A.e · Hughes C.V.f · Pradhan N.g · Loo C.Y.g · Tanner A.C.R.a, d
aDepartment of Molecular Genetics and bDepartment of Biostatistics, The Forsyth Institute, cOral Health Policy and Epidemiology, dDepartment of Oral Medicine, Infection and Immunity and eHarvard School of Dental Medicine, Harvard University, fDepartment of Pediatric Dentistry, Goldman School of Dental Medicine, Boston University, and gDepartment of Pediatric Dentistry, Tufts School of Dental Medicine, Tufts University, Boston, Mass., USA Corresponding Author
Anne C.R. Tanner
The Forsyth Institute
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Boston MA 02115 (USA)
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Background/Aims: Severe early childhood caries is a microbial infection that severely compromises the dentition of young children. The aim of this study was to characterize the microbiota of severe early childhood caries. Methods: Dental plaque samples from 2- to 6-year-old children were analyzed using 16S rRNA gene cloning and sequencing, and by specific PCR amplification for Streptococcus mutans and Bifidobacteriaceae species. Results: Children with severe caries (n = 39) had more dental plaque and gingival inflammation than caries-free children (n = 41). Analysis of phylotypes from operational taxonomic unit analysis of 16S rRNA clonal metalibraries from severe caries and caries-free children indicated that while libraries differed significantly (p < 0.0001), there was increased diversity than detected in this clonal analysis. Using the Human Oral Microbiome Database, 139 different taxa were identified. Within the limits of this study, caries-associated taxa included Granulicatella elegans (p < 0.01) and Veillonella sp. HOT-780 (p < 0.01). The species associated with caries-free children included Capnocytophaga gingivalis (p < 0.01), Abiotrophia defectiva (p < 0.01), Lachnospiraceae sp. HOT-100 (p < 0.05), Streptococcus sanguinis (p < 0.05) and Streptococcus cristatus (p < 0.05). By specific PCR, S. mutans (p < 0.005) and Bifidobacteriaceae spp. (p < 0.0001) were significantly associated with severe caries. Conclusion: Clonal analysis of 80 children identified a diverse microbiota that differed between severe caries and caries-free children, but the association of S. mutans with caries was from specific PCR analysis, not from clonal analysis, of samples.
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