口腔需氧菌及兼性厌氧菌种分离鉴定系统的建立  

Isolation and identification of aerobic and facultative anaerobic bacteria in the oral cavity

作　　者：陆文昕 吴凡子 周辛璇 吴岚[2] 李明云[1] 任彪[1] 郭强[1] 黄睿洁 李继遥[1] 肖丽英[1] 李燕[1] 

机构地区：[1]口腔疾病研究国家重点实验室//华西口腔医学院//四川大学,四川成都610041 [2]四川农业大学资源学院微生物学,四川成都611130

出　　处：《南方医科大学学报》2015年第12期1710-1714,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(81430011);国家科技支撑计划项目(2012BAI07B00);四川省科技厅科技支撑计划项目(2013SZ0039)~~

摘　　要：目的建立口腔需氧菌和兼性菌的系统分离鉴定方法。方法招募20名口腔健康志愿者及8名口腔疾病患者,分别采集唾液、龈下菌斑、龈上菌斑和根尖周肉芽样本,采用形态鉴定、全自动生化鉴定仪及16s r RNA基因测序的多相分类方法,对口腔可培养细菌进行分离鉴定。结果鉴定出口腔菌种63种,共175株,梅里埃生化鉴定与16s r RNA基因鉴定的符合度为22.39%,不符合的菌种以基因测序结果为准。链球菌属、放线菌属和葡萄球菌属检出率占前3位,菌种以咽峡炎链球菌、口腔放线菌、变异链球菌、缓症链球菌检出频率最高。慢性根尖周炎的咽峡炎链球菌检出率最高;放射性龋患者中,中间链球菌、金黄色葡萄球菌检出率较高;猛性龋患者中,变异链球菌远高于其他细菌。结论口腔需氧菌和兼性厌氧菌中以革兰阳性菌检出率最高。对于口腔细菌鉴定来说,梅里埃生化鉴定仪鉴定准确率不高,但可提供菌株生理生化指标,而全长的16s r RNA基因测序鉴定更为可靠。Objective To establish a systematic method for isolation and identification of aerobic and facultative anaerobic bacteria in the oral cavity. Methods Samples of the saliva, dental plaque and periapical granulation tissue were collected from20 subjects with healthy oral condition and from 8 patients with different oral diseases. The bacteria in the samples were identified by morphological identification, VITEK automatic microorganism identification and 16 s r RNA gene sequencing.Results VITEK automatic microorganism identification and 16 s r RNA gene sequencing showed an agreement rate of 22.39% in identifying the bacteria in the samples. We identified altogether 63 bacterial genus（175 species）, among which Streptococcus,Actinomyces and Staphylococcus were the most common bacterial genus, and Streptococcus anginosus, Actinomyces oris,Streptococcus mutans and Streptococcus mitis were the most common species. Streptococcus anginosus was commonly found in patients with chronic periapical periodontitis. Streptococcus intermedius and Staphylococcus aureus were common in patients with radiation caries, and in patients with rampant caries, Streptococcus mutans was found at considerably higher rate than other species. Conclusion Aerobic and facultative anaerobic bacteria are commonly found in the oral cavity, and most of them are gram-positive. 16 s r RNA gene sequencing is more accurate than VITEK automatic microorganism identification in identifying the bacteria.

关 键 词：生化鉴定 16s r RNA基因测序 菌种鉴定系统 

分 类 号：R446.5[医药卫生—诊断学]