持续性根尖周炎中牙龈卟啉单胞菌的分离与鉴定  

Isolation and identification of porphyromonas gingivalis clinical strains from persistent periodontitis lesions

作　　者：姚雪芹[1,2] 侯本祥[1] 张琛[3] 

机构地区：[1]首都医科大学附属北京口腔医院牙体牙髓科,硕士生北京100050 [2]北京市通州区新华医院口腔综合科,主治医师北京101100 [3]首都医科大学北京口腔医院牙体牙髓科,副主任医师副教授北京100050

出　　处：《中华老年口腔医学杂志》2016年第2期77-82,共6页Chinese Journal of Geriatric Dentistry

基　　金：国家自然科学基金(项目编号:81200783);北京市卫生系统高层次卫生技术人才培养计划(项目编号:2013-3-039)

摘　　要：目的:建立持续性根尖周炎中牙龈卟啉单胞菌的分离鉴定方法。方法:选择持续性根尖周炎患者5例,通过严格无菌的根尖手术,从切除的根尖及根尖周组织中获得临床标本,厌氧分离提纯培养,通过形态学观察,生化反应鉴定及分子生物学16Sr RNA特异性片段扩增测序,得到牙龈卟啉单胞菌临床分离菌株。结果:5例临床样本得到4株培养墨黑色菌落,表面似金属光泽,完全溶血。革兰染色为阴性球杆菌。生化反应符合牙龈卟啉单胞菌生化特点,琼脂糖凝胶电泳得到目标条带,4株临床菌株16Sr RNA基因扩增测序,与NCBI数据库对比,得到一株临床菌株与牙龈卟啉单胞菌16Sr RNA基因相似度大于99.6%,其余3株为中间普雷沃菌。结论:本研究严格选取临床样本,形态学方法、生化学方法以及分子生物学鉴定方法可分离并严格有效鉴定分离菌株,从持续性根尖周炎中分离获得牙龈卟啉单胞菌临床株,为研究持续性根尖周炎中牙龈卟啉单胞菌的致病机制奠定基础。Objective： To establish isolation and identification method of Porphyromonas gingivalis from the persistent periodontitis lesions. Methods： Five apex samples and apical tissue were obtained directly from apical lesions of persistent root periodontitis during root-end surgery.All procedures were performed in sterile manner to avoid bacterial contamination.Isolated strain were detected by microbial morphology, biochemistry method, 16 Sr RNA gene sequencing and specific PCR amplification. Results：Four strains of ink black colonies with metal luster, gram negative baillus, were obtained from 5 clinical samples. The strains were characteristics consistent with the biochemical charcteristics of Porphyromonas gingivalis. By 16 Sr RNA sequencing and conducting BLAST in NCBI detabase, one isolated strain owned 99.6%similarity with Porphyromonas gingivalis sequences and had the specific band in PCR amplification. Conclusion： A strain of porphyromonas gingivalis strain isolated from persistent apical periodontitis sample was identified by morphology,biochemistry and molecular biology method.

关 键 词：持续性根尖周炎 牙龈卟啉单胞菌 细菌分离与鉴定 

分 类 号：R781.4[医药卫生—口腔医学]