Molecular and biological classification of periodontopathic spirochetes

牙周病螺旋体的分子和生物学分类

• 批准号: 09671879
• 负责人: UMEMOTO Toshihiko
• 金额: $ 1.66万
• 依托单位: Asahi University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671879/
• 关键词: Oral spirochetes; Treponema medium; PCR analysis; Periodontal disease; Surface proteins; 16S rRNA gene; phylogenic analysis; Coaggregation

Some human oral treponemes have been associated with soft-tissue damages, and eventually these lead to periodontal tissue destruction in certain types of periodontal disease. We planed to distinguish these diverse microorganisms by molecular and biological characteristics.1. We examined the phenotypic, serologic, and genetic characteristics of an isolated medium-sized treponeme, compared the results with data for reference species, and proposed a new species, Treponema medium. Its GC content was 51 mole%. The DNA-DNA relatedness of this organism to other Treponema species were less than 30%, suggesting that this microorganism is genetically distinct from the established oral Treponema species.2. Type I-, IV-, and V-collagen-binding polypeptides (CBPs) were detected in the preparations from both T.denticola and T.socranskii subsp. buccale. Few CBPs, however, were detected in the preparations from T.medium. Immunoelectron microscopy using rabbit antisera against the CBPs demonstrated tha … More t the CBPs were located on the outer envelopes. The adherence of T.denticola to the collagen-coated surfaces was significantly greater than that of T.medium.3. PCR assay using primers specific for the genomic DNA segment coding 165 rRNA of T.medium was examined for detecting the organism in subgingival plaque samples from patients with periodontitis. The PCR analysis resulted in a single band of the PCR products. The PCR assay detected 10 to 100 T.medium cells. The primers reacted with both T.medium and T.vincentii, while neither of the primers reacted with other oral or non-oral bacterial species. The PCR analysis showed a definite relativity in subgingival plaque samples of patients with moderate to severe periodontitis.4. T.medium strongly coaggregated with fimbriate P.gin givalis strains, but not with afimbriate strains. Heated P.gingivalis cells lost their ability to bind the heated or unheated T.medium cells. The antiserum against the fimbriae of P.gingivalis ATCC 33277 produced different "aggregates' consisting of P.gingivalis cells with few spirochetes, but the preimmune serum did not disturb the coaggregation reaction. A heat-stable 37kDa surface protein of T.medium attached to the P.gingivalis fimbriae. This is the first report of fimbriae-mediated coaggregation between human oral spirochetes and P.gingivalis. Less

一些人类口腔密螺旋体与软组织损伤有关，并最终导致某些类型牙周病的牙周组织破坏。我们计划通过分子和生物学特性来区分这些不同的微生物。我们检测了一个分离的中型密螺旋体的表型、血清学和遗传特征，并将结果与参考物种的数据进行了比较，提出了一个新种——密螺旋体培养基。其GC含量为51摩尔%。该微生物与其他密螺旋体物种的DNA-DNA亲缘性小于30%，表明该微生物与已建立的口腔密螺旋体物种具有遗传差异。在齿状棘球蚴和索氏棘球蚴的制剂中均检测到I型、IV型和v型胶原结合多肽（CBPs）。buccale。然而，在t培养基的制备中检测到很少的CBPs。利用兔抗血清对CBPs进行免疫电镜观察，发现CBPs主要位于外包膜上。齿状棘球蚴对胶原膜表面的粘附力明显大于中棘球蚴。采用引物对T.medium编码165 rRNA的基因组DNA片段进行PCR检测，用于检测牙周炎患者龈下菌斑样本中的T.medium。PCR分析结果为单带PCR产物。PCR检测到10 ~ 100个T.medium细胞。引物与T.medium和t .vincent均有反应，而与其他口腔或非口腔细菌均无反应。PCR分析显示，中重度牙周炎患者的龈下菌斑样本具有一定的相关性。T.medium与非拟合菌强共聚，与非拟合菌不强共聚。加热后的牙龈假单胞菌细胞失去了与加热或未加热的假单胞菌细胞结合的能力。针对牙龈卟啉菌ATCC 33277菌毛的抗血清产生由牙龈卟啉菌细胞和少量螺旋体组成的不同“聚集体”，但免疫前血清不干扰共聚集体反应。附着在牙龈假单胞菌菌毛上的一种热稳定的37kDa的t .培养基表面蛋白。这是首次报道人类口腔螺旋体与牙龈卟啉单胞菌通过菌毛介导的共聚集。少

项目成果

Nakazawa, F., Umemoto, T., Namikawa, I., Hoshino, E.: "Genetic characterization of an oral treponeme isolated from human subgingival plaque." Oral Microbiol.Immunol.12(3). 189-192 (1997)

Nakazawa, F.、Umemoto, T.、Namikawa, I.、Hoshino, E.：“从人类龈下菌斑中分离出的口腔密螺旋体的遗传特征。”

梅本利彦: "ヒト口腔スピロヘータ研究の最前線" 日本細菌学雑誌. 53(2). 425-433 (1998)

Toshihiko Umemoto：“人类口腔螺旋体研究的前沿”日本细菌学杂志 53(2) 425-433 (1998)。

Umemoto, T., Nakazawa, F., Hoshino, E., Okada, K., Fukunaga, M., Namikawa, I.: "Treponema medium sp.nov., isolated from human subgingival dental plaque." Int.J.Syst. Bacteriol.47(1). 67-72 (1997)

Umemoto, T.、Nakazawa, F.、Hoshino, E.、Okada, K.、Fukunaga, M.、Namikawa, I.：“Treponema media sp.nov.，从人类龈下牙菌斑中分离出来。”

Kusumoto,Y.,Yamamoto,H.,Kureshiro,H.,Umemoto,T.,Ogawa,T.: "Oral spirochetal species detection using a polymerase chain reaction." Dentistry in Japan. 35(in press). (1999)

Kusumoto,Y.、Yamamoto,H.、Kureshiro,H.、Umemoto,T.、Okawa,T.：“使用聚合酶链反应检测口腔螺旋体物种。”

Umemoto, T., Li, M., Namikawa, I.: "Adherence of human oral spirochetes by collagen-binding proteins." Microbiol.Immunol.41(12). 917-923 (1997)

Umemoto, T.、Li, M.、Namikawa, I.：“胶原蛋白结合蛋白对人类口腔螺旋体的粘附。”