Aas Jørn A, Griffen Ann L, Dardis Sara R, Lee Alice M, Olsen Ingar, Dewhirst Floyd E, Leys Eugene J, Paster Bruce J
Institute of Oral Biology, University of Oslo, Post Box 1052 Blindern, 0316 Oslo, Norway.
J Clin Microbiol. 2008 Apr;46(4):1407-17. doi: 10.1128/JCM.01410-07. Epub 2008 Jan 23.
Although Streptococcus mutans has been implicated as a major etiological agent of dental caries, our cross-sectional preliminary study indicated that 10% of subjects with rampant caries in permanent teeth do not have detectable levels of S. mutans. Our aims were to use molecular methods to detect all bacterial species associated with caries in primary and permanent teeth and to determine the bacterial profiles associated with different disease states. Plaque was collected from 39 healthy controls and from intact enamel and white-spot lesions, dentin lesions, and deep-dentin lesions in each of 51 subjects with severe caries. 16S rRNA genes were PCR amplified, cloned, and sequenced to determine species identities. In a reverse-capture checkerboard assay, 243 samples were analyzed for 110 prevalent bacterial species. A sequencing analysis of 1,285 16S rRNA clones detected 197 bacterial species/phylotypes, of which 50% were not cultivable. Twenty-two new phylotypes were identified. PROC MIXED tests revealed health- and disease-associated species. In subjects with S. mutans, additional species, e.g., species of the genera Atopobium, Propionibacterium, and Lactobacillus, were present at significantly higher levels than those of S. mutans. Lactobacillus spp., Bifidobacterium dentium, and low-pH non-S. mutans streptococci were predominant in subjects with no detectable S. mutans. Actinomyces spp. and non-S. mutans streptococci were predominant in white-spot lesions, while known acid producers were found at their highest levels later in disease. Bacterial profiles change with disease states and differ between primary and secondary dentitions. Bacterial species other than S. mutans, e.g., species of the genera Veillonella, Lactobacillus, Bifidobacterium, and Propionibacterium, low-pH non-S. mutans streptococci, Actinomyces spp., and Atopobium spp., likely play important roles in caries progression.
虽然变形链球菌一直被认为是龋齿的主要病因,但我们的横断面初步研究表明,10% 的恒牙患猖獗龋的受试者检测不到变形链球菌。我们的目的是使用分子方法检测与乳牙和恒牙龋齿相关的所有细菌种类,并确定与不同疾病状态相关的细菌谱。从39名健康对照者以及51名患有严重龋齿的受试者的完整牙釉质、白斑病变、牙本质病变和深牙本质病变处采集菌斑。对16S rRNA基因进行PCR扩增、克隆和测序以确定物种身份。在反向捕获棋盘分析中,对243个样本进行了110种常见细菌的分析。对1285个16S rRNA克隆的测序分析检测到197种细菌种类/系统发育型,其中50% 不可培养。鉴定出22种新的系统发育型。PROC MIXED检验揭示了与健康和疾病相关的物种。在携带变形链球菌的受试者中,其他物种,如阿托波氏菌属、丙酸杆菌属和乳杆菌属的物种,其含量明显高于变形链球菌。在检测不到变形链球菌的受试者中,乳杆菌属、龋齿双歧杆菌和低pH值非变形链球菌占主导地位。放线菌属和非变形链球菌在白斑病变中占主导地位,而已知的产酸菌在疾病后期含量最高。细菌谱随疾病状态而变化,在乳牙列和恒牙列之间也有所不同。除变形链球菌外的其他细菌种类,如韦荣氏菌属、乳杆菌属、双歧杆菌属、丙酸杆菌属、低pH值非变形链球菌、放线菌属和阿托波氏菌属的物种,可能在龋齿进展中起重要作用。