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目的分析患者口腔全微生物种类,对猖獗龋患者提出细菌学预防与治疗手段。方法抽血查免疫全套和血常规,获取全菌斑(A)、龋损(Q)、唾液(T)、黏膜(N)样本经原代涂片镜检,并传代分离培养后采用形态学、梅里埃鉴定仪、16S r DNA序列进化分析鉴定微生物种类;进一步采用Q-PCR分析变异链球菌的含量,DGGE分析微生物组成和结构。结果血液免疫全套基本正常,但Ig E、中性分叶核细胞增高。原代样本涂片发现唾液中有大量极长链的细菌,经培养发现唾液中总菌含量最高,龋损样本中溶血菌含量最高,变异链球菌在全菌斑中最多。梅里埃和测序鉴定出33株细菌,全菌斑中分离出多种链球菌和韦德纤毛菌、龋损菌斑中多次分离到变异链球菌、具核梭杆菌和苏黎世链球菌,唾液样本中分离出多种乳杆菌。q PCR发现变异链球菌含量明显增高;变性梯度凝胶电泳(DGGE)显示该患者细菌多样性明显降低,但丰度明显增强;条带切胶测序检出多种纤毛菌。结论患者免疫功能基本正常,血液检测呈现感染血象,病因疑与细菌感染有关。可能由于唾液减少导致变异链球菌、多种纤毛菌等致病微生物过度繁殖、细菌多样性减少而引起生态失衡。
Objective To analyze the oral microbiota in patients with bacterial caries prevention and treatment of rampant means. Methods A total of blood samples were collected for blood tests and blood tests were performed. Samples of whole plaque (A), carious lesions (Q), saliva (T) and mucosa (N) were obtained by primary smear microscopy and subculture. , Merier identification instrument, 16S r DNA sequence evolution analysis to identify microbial species; further using Q-PCR analysis of Streptococcus mutans content, DGGE analysis of microbial composition and structure. Results The whole set of blood immunization was basically normal, but Ig E and neutral lobule nuclear cells increased. The primary sample smear found a large number of very long chain bacteria in saliva. The highest content of total bacteria in saliva was found after culture, the highest content of hemolytic bacteria in the carious samples, and the most streptococcus mutans in the whole plaque. Meridian and sequencing identified 33 strains of bacteria, streptococcus and Wade bacteria were isolated from plaque, and S. mutans and B. liquefiens were isolated from cariogenic plaque. Salivary samples In a variety of isolated Lactobacillus. q PCR showed that the content of Streptococcus mutans was significantly increased; denaturing gradient gel electrophoresis (DGGE) showed that the bacterial diversity was significantly decreased, but the abundance was significantly enhanced; Conclusions The immune function of patients is basically normal, the blood test shows the blood of infection, and the cause of the disease is suspected to be related to bacterial infection. May be due to reduced saliva led to Streptococcus mutans, a variety of cilia and other pathogenic microorganisms over-breeding, reduce bacterial diversity and cause ecological imbalance.