慢性牙周炎龈下菌斑中福赛斯坦纳菌的检测

Quantification of Tannerella forsythensis in chronic periodontitis patients

目的检测慢性牙周炎患者和牙周健康者龈下菌斑中福赛斯坦纳菌的数量和所占比例,探讨福赛斯坦纳菌与牙周炎发生发展的关系。方法采集经常规聚合酶链反应(PCR)法检测福赛斯坦纳菌为阳性的61例慢性牙周炎患者和12例牙周健康者的龈下菌斑,应用TaqMan实时荧光定量PCR法对菌斑的细菌总量和福赛斯坦纳菌的数量进行定量检测,构建含有福赛斯坦纳菌和真细菌靶基因的重组质粒,建立定量标准。结果本研究设计的引物和探针具有良好的特异性和敏感性;慢性牙周炎患者病变位点的福赛斯坦纳菌数量和细菌总量均高于牙周健康者的健康位点,且福赛斯坦纳菌在龈下菌斑中的比例也比健康位点高(P<0.05);龈下菌斑的细菌数量与探诊深度呈正相关(P<0.001);龈下菌斑中福赛斯坦纳菌所占比例在不同的探诊深度间无统计学差异(P>0.05)。结论龈下菌斑中福赛斯坦纳菌的数量与牙周状况有密切关系,实时荧光定量PCR法是研究牙周病病因及治疗方法的有效手段。

Objective To compare the number of the Tannerella forsythensis （T forsythensis）, total bacteria, and proportion of T. forsythensis in subgingival specimens in diseased sites of chronic periodontitis patients and in healthy sites of periodontally healthy subjects, and clarify the relationship between bacterial load and periodontal status. Methods Subgingival plaque samples from 61 chronic periodontitis patients and 12 healthy controls（positive for T. forsythensis by conventional PCR） were analyzed with TaqMan real-time polymerase chain reaction assay for T. forsythensis and total bacteria. Quantification was performed with species-specific primer/probe, universal primer/ probe and serial dilution of plasmid standards. Results Numbers of T. forsythensis and total bacteria（P〈0.001） , the proportion of T. forsythensis in subgingival specimens （P〈0.05）were significant higher in diseased sites of chronic periodontitis patients than in healthy sites of healthy subjects. In addition, a significant correlation was found between the number of bacteria and various probing depth （P〈0.001）. There was no significantly difference between the proportion of T. forsythensis in subgingival plaque and probing depth. Conclusion Number of T. forsythensis are closely associated with periodontal status, and demonstrate the broad potential of real-time polymerase chain reaction application on periodontology.