Detection and quantification of oral treponemes in subgingival plaque by real-time PCR
Detection and quantification of oral treponemes in subgingival plaque by real-time PCR
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DOI：
10.1128/jcm.40.9.3334-3340.2002
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发表时间：
2002-09-01
影响因子：
9.4
通讯作者：
Ogawa, T
中科院分区：
文献类型：
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作者：
Asai, Y;Jinno, T;Ogawa, T
口腔密螺旋体与牙周病有关。我们开发了一种高度敏感和特异性的方法来检测和定量可培养的口腔密螺旋体（齿垢密螺旋体，Vincentii密螺旋体和密螺旋体培养基）在50龈下菌斑样本从13名健康受试者以及37例牙周病患者使用实时PCR检测与特定引物和TaqMan探针的每个16 S rRNA序列。通过使用来自各种密螺旋体和其他细菌物种的DNA样本检查每个检测的特异性。TaqMan实时PCR能够检测10（3）至108个口腔密螺旋体细胞，相关系数如下：T。denticola，0.984; T. vincentii，0.991; T.中等，0.984。龈下菌斑样本中这三种口腔密螺旋体的出现频率如下：T。齿垢类68.0%; T. vincentii，36.0%;中等，48.0%。此外，T. denticola，齿齿叶蜂T. vincentii和T.通过实时PCR检测的空斑样品中的中等细胞的范围分别为3至15，184、1至447和1至7，301个细胞/pg空斑DNA。增加了T。在深层牙周袋的牙菌斑样本中检测到齿垢细胞，并且T.在深口袋中也发现了中等。另一方面，T. Vincentii主要分布在较浅的口袋中。这些结果表明，各种口腔密螺旋体与牙周病的各个阶段的形成。
Oral treponemes have been associated with periodontal diseases. We developed a highly sensitive and specific method to detect and quantify cultivable oral treponemes (Treponema denticola, Treponema vincentii, and Treponema medium) in 50 subgingival plaque samples from 13 healthy subjects as well as 37 patients with periodontal diseases using real-time PCR assays with specific primers and a TaqMan probe for each 16S rRNA sequence. The specificity for each assay was examined by using DNA specimens from various treponemal and other bacterial species. The TaqMan real-time PCR was able to detect from 10(3) to 108 cells of the oral treponemes, with correlation coefficients as follows: T. denticola, 0.984; T. vincentii, 0.991; and T. medium, 0.984. The frequencies of occurrence of these three oral treponemes in subgingival plaque samples were as follows: T. denticola, 68.0%; T. vincentii, 36.0%; and T. medium, 48.0%. In addition, the number of T. denticola, T. vincentii, and T. medium cells in plaque samples detected by real-time PCR ranged from 3 to 15,184, 1 to 447, and I to 7,301 cells/pg of plaque DNA, respectively. Increased numbers of T. denticola cells were detected in plaque samples from deep periodontal pockets, and T. medium was also detected in deep pockets. On the other hand, T. vincentii was mainly found in shallow pockets. These results suggest that various oral treponemes are associated with the formation of each stage of periodontal disease.