Quantitative analysis of diverse Lactobacillus species present in advanced dental caries
Quantitative analysis of diverse Lactobacillus species present in advanced dental caries
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DOI：
10.1128/jcm.42.7.3128-3136.2004
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发表时间：
2004-07-01
影响因子：
9.4
通讯作者：
Hunter, N
中科院分区：
文献类型：
--
作者：
Byun, R;Nadkarni, MA;Hunter, N
我们以前的分析65个先进的龋齿病变的传统培养技术表明，乳酸杆菌是众多的前进前沿的进行性病变。乳酸杆菌产生的有机酸被认为是导致牙本质基质脱钙的重要因素。本研究进行更精确地定义在这种环境中发现的乳酸杆菌的多样性，并通过实时PCR定量相对于乳酸杆菌的总负荷的主要物种和乳酸杆菌类型。合并的DNA通过PCR用乳杆菌属特异性引物扩增，用于随后的克隆、测序和系统发育分析。基于16S核糖体DNA序列比较，检测到18种不同的乳酸杆菌，包括新的和胃肠道乳酸杆菌类型的强代表性。针对加氏乳杆菌（Lactobacillus gasseri）、L. ultunensis，L.唾液腺L.鼠李属L.干酪湖（L.casei）crispatus，L.德尔布吕茨基湖fermentum和L.鸡。在大多数牙本质样本中发现了三种以上的不同菌种，证实了各种乳杆菌的广泛分布和数量重要性。在龋性牙本质中。实时PCR定量显示，在龋齿牙本质上定植的九种物种的比例各不相同，其中L. gasseri和L. ultunensis比其他流行的物种。研究结果为进一步研究乳酸杆菌的致病性奠定了基础。在龋损扩大的情况下。
Our previous analysis of 65 advanced dental caries lesions by traditional culture techniques indicated that lactobacilli were numerous in the advancing front of the progressive lesion. Production of organic acids by lactobacilli is considered to be important in causing decalcification of the dentinal matrix. The present study was undertaken to define more precisely the diversity of lactobacilli found in this environment and to quantify the major species and phylotypes relative to total load of lactobacilli by real-time PCR. Pooled DNA was amplified by PCR with Lactobacillus genus-specific primers for subsequent cloning, sequencing, and phylogenetic analysis. Based on 16S ribosomal DNA sequence comparisons, 18 different phylotypes of lactobacilli were detected, including strong representation of both novel and gastrointestinal phylotypes. Specific PCR primers were designed for nine prominent species, including Lactobacillus gasseri, L. ultunensis, L. salivarius, L. rhamnosus, L. casei, L. crispatus, L. delbrueckii, L. fermentum, and L. gallinarum. More than three different species were identified as being present in most of the dentine samples, confirming the widespread distribution and numerical importance of various Lactobacillus spp. in carious dentine. Quantification by real-time PCR revealed various proportions of the nine species colonizing carious dentine, with higher mean loads of L. gasseri and L. ultunensis than of the other prevalent species. The findings provide a basis for further characterization of the pathogenicity of Lactobacillus spp. in the context of extension of the carious lesion.