Sakamoto Mitsuo, Huang Yi, Umeda Makoto, Ishikawa Isao, Benno Yoshimi
Japan Collection of Microorganisms, RIKEN, Wako, 351-0198, Saitama, Japan.
FEMS Microbiol Lett. 2002 Nov 19;217(1):65-9. doi: 10.1111/j.1574-6968.2002.tb11457.x.
A phylogenetic approach based on 16S rRNA (rDNA) has been recently applied to investigate the diversity of cultivable and uncultivable species in the human oral cavity without cultivation. In a previous study [Sakamoto et al. (2000) Microbiol. Immunol. 44, 643-652], we identified a number of novel oral phylotypes, representing as yet uncultured organisms. The purpose of this study was to design specific PCR primers for five phylotypes AP12, AP21, AP24, AP50, and RP58, which are deeply branched particularly in the phylogenetic tree, and determine the prevalence of these phylotypes in 45 patients with periodontitis and 18 healthy subjects. The specificity of each primer was validated by the sequence analysis of PCR products obtained from saliva and subgingival plaque samples. Among phylotypes tested, phylotype AP24, which is closely related to oral clone DA014 reported previously [Paster et al. (2001) J. Bacteriol. 183, 3770-3783], was significantly associated with saliva and subgingival plaque samples from patients with periodontitis (P<0.01), but the difference was not statistically significant in the presence of other phylotypes. These data suggest that phylotype AP24 may play an important role in periodontal disease.
一种基于16S rRNA（rDNA）的系统发育方法最近被应用于在不进行培养的情况下研究人类口腔中可培养和不可培养物种的多样性。在之前的一项研究中[Sakamoto等人（2000年），《微生物学与免疫学》44卷，643 - 652页]，我们鉴定出了许多新型口腔系统发育型，它们代表了尚未培养的生物体。本研究的目的是为五个系统发育型AP12、AP21、AP24、AP50和RP58设计特异性PCR引物，这些系统发育型在系统发育树中分支特别深，并确定这些系统发育型在45名牙周炎患者和18名健康受试者中的流行情况。通过对从唾液和龈下菌斑样本中获得的PCR产物进行序列分析，验证了每个引物的特异性。在所测试的系统发育型中，与先前报道的口腔克隆DA014密切相关的系统发育型AP24[Paster等人（2001年），《细菌学杂志》183卷，3770 - 3783页]，与牙周炎患者的唾液和龈下菌斑样本显著相关（P<0.01），但在存在其他系统发育型的情况下，差异无统计学意义。这些数据表明系统发育型AP24可能在牙周疾病中起重要作用。
