Cultivable Anaerobic Microbiota of Severe Early Childhood Caries
Cultivable Anaerobic Microbiota of Severe Early Childhood Caries
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DOI：
10.1128/jcm.02427-10
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发表时间：
2011-04-01
影响因子：
9.4
通讯作者：
Dewhirst, F. E.
中科院分区：
文献类型：
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作者：
Tanner, A. C. R.;Mathney, J. M. J.;Dewhirst, F. E.
严重的儿童早期龋齿 (ECC) 虽然通过选择性检测（培养、PCR）与变形链球菌密切相关，但通过分子克隆方法也与广泛多样化的微生物群相关。本研究的目的是利用厌氧培养评估严重 ECC 的微生物群。将 42 名严重 ECC 儿童的牙菌斑微生物组成与 40 名无龋儿童的牙菌斑微生物组成进行比较。细菌样品在血液和酸性（pH 5）琼脂上进行厌氧培养。对分离株进行纯化，并从 5,608 个分离株中获得了 16S rRNA 基因的部分序列。对来自严重 ECC 和无龋儿童的血液和酸琼脂的 16S rRNA 分离物文库进行基于序列的分析，其人群覆盖率 > 90%，并且血液分离物文库中存在更大的多样性。将分离序列与人类口腔微生物组数据库 (HOMD) 中的分类单元序列进行比较，鉴定出 198 个 HOMD 分类单元，其中包括 45 个以前未培养的分类单元、29 个扩展的 HOMD 分类单元和 45 个潜在的新类群。与严重 ECC 相关的主要物种包括变形链球菌、Scardovia wiggsiae、小韦荣球菌、冠状链球菌和放线菌。在存在或不存在变形链球菌检测的情况下，维氏链球菌与严重 ECC 儿童显着相关。我们得出的结论是，厌氧培养在 ECC 中检测到的物种多样性与使用克隆方法观察到的物种多样性一样广泛。培养与 16S rRNA 鉴定相结合，鉴定出超过 74 个人类口腔分类群的分离株，而此前没有培养出代表。主要的龋齿相关物种是变形链球菌和威格斯链球菌，后者是新认识的龋齿病原体的候选者。
Severe early childhood caries (ECC), while strongly associated with Streptococcus mutans using selective detection (culture, PCR), has also been associated with a widely diverse microbiota using molecular cloning approaches. The aim of this study was to evaluate the microbiota of severe ECC using anaerobic culture. The microbial composition of dental plaque from 42 severe ECC children was compared with that of 40 caries-free children. Bacterial samples were cultured anaerobically on blood and acid (pH 5) agars. Isolates were purified, and partial sequences for the 16S rRNA gene were obtained from 5,608 isolates. Sequence-based analysis of the 16S rRNA isolate libraries from blood and acid agars of severe ECC and caries-free children had >90% population coverage, with greater diversity occurring in the blood isolate library. Isolate sequences were compared with taxon sequences in the Human Oral Microbiome Database (HOMD), and 198 HOMD taxa were identified, including 45 previously uncultivated taxa, 29 extended HOMD taxa, and 45 potential novel groups. The major species associated with severe ECC included Streptococcus mutans, Scardovia wiggsiae, Veillonella parvula, Streptococcus cristatus, and Actinomyces gerensceriae. S. wiggsiae was significantly associated with severe ECC children in the presence and absence of S. mutans detection. We conclude that anaerobic culture detected as wide a diversity of species in ECC as that observed using cloning approaches. Culture coupled with 16S rRNA identification identified over 74 isolates for human oral taxa without previously cultivated representatives. The major caries-associated species were S. mutans and S. wiggsiae, the latter of which is a candidate as a newly recognized caries pathogen.