维吾尔族老年慢性牙周炎患者唾液中福塞斯坦纳菌的检测

Detection of tannerella forsythensis in saliva of older Uigur patients with chronic periodontitis

目的应用聚合酶链反应(polymerase chain reaction,PCR)检测维吾尔族老年慢性牙周炎患者唾液及龈下菌斑中福赛斯坦纳菌(Tannerella forsythensis,T.f),并探讨该菌在唾液标本与龈下菌斑标本检测结果的一致性。方法选择新疆和田地区维吾尔族老年慢性牙周炎患者71例。采集唾液及龈下菌斑标本,使用浓度25%的Chelex-100(一种螯合型树脂)制备DNA模板。应用T.f的16srRNA引物,分别对唾液及龈下菌斑标本进行PCR扩增,部分扩增产物克隆后测定核苷酸序列,并观察T.f的检出率,计算Kappa值。结果唾液和龈下菌斑标本中T.f的检测结果具有高度一致性,其检出率分别为76.06%(54/71)和90.14%(64/71),Kappa值为0.322,一致率为80.28%。T.f的16SrRNA基因片段克隆测序结果与Blast比对,与Genebank中已登记的16srRNA基因片段同源性为100%。结论 (1)采用唾液标本检测T.f的灵敏度高且取材方便,有可能取代龈下菌斑标本。(2)新疆和田地区维吾尔族老人慢性牙周炎患者T.f检出率较高。

Objective To detect Tannerella forsythensis （T.f） in saliva of older Uigur patients with chronic periodontitis and compare the test results of T.f from saliva and subgingival plaque. Methods Subgingival plaque and saliva were collected from 71 older Uigur patients with chronic periodontitis in Hetian. Subgingival plaque and saliva treated with 25% chelex-100（a cation-chelating resin）to prepare DNA templates. The fragments of 16s rRNA gene were detected by PCR with a pair of primers in T.f,and sequenced. The detection rate of T.f was calculated and the kappa test used as a chance-corrected measure of agreement between saliva and pocket samples.Results T.f was detected in 76.06%（54/71）of saliva and 90.14%（64/71）of subgingival plaque sample.The kappa test value was 0.322,with a accurate rate of 80.28%. Sequencing results of 16s rRNA gene fagments demonstrated that it had 100% similarity with reported at Genebank. Conclusion （1） Saliva was a good substitute for subgingival plaque for determining the presence of T.f in mouth.（2） There is a high detection rate of T.f in older Uigur patients with chronic periodontitis in Hetian was calculated.