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目的分离鉴定龋病患儿口腔中双歧杆菌的存在情况,探讨口腔双歧杆菌与儿童龋病的相关性。方法选取29例3~6岁儿童为研究对象,其中实验组(重症龋病)16例,对照组(无龋病)13例。采集两组儿童龈上的混合菌斑和非刺激性唾液,采用改良的双歧杆菌选择性培养基进行分离培养,对培养后形成的单菌落采用双歧杆菌特异性引物进行分子生物学鉴定。结果分离培养后实验组儿童菌斑和唾液样本中双歧杆菌的检出率均明显高于对照组,差异有统计学意义(P<0.05)。从实验组检出双歧杆菌的样本中随机选取单菌落进行PCR扩增后测序,结果显示实验组菌斑60个菌落中齿双歧杆菌占81.67%、栖牙双歧杆菌占6.67%、殊形双歧杆菌占11.66%;唾液样本60个菌落中齿双歧杆菌占88.33%、栖牙双歧杆菌占3.34%、殊形双歧杆菌占8.33%。结论双歧杆菌与儿童重症龋病的发生有一定相关性,儿童口腔双歧杆菌中主要菌株为齿双歧杆菌、栖牙双歧杆菌、殊形双歧杆菌,其中齿双歧杆菌比例最高,是儿童龋病的活跃菌种。
Objective To isolate and identify the presence of Bifidobacterium in oral cavity of children with dental caries and to explore the correlation between Bifidobacterium oralis and caries in children. Methods Twenty-nine children aged 3 to 6 years old were enrolled in the study, including 16 cases of severe caries in experimental group and 13 cases of control group (caries-free). The mixed plaque and non-irritating saliva of the two groups of children were collected. The modified Bifidobacterium selective medium was used for isolation and culture. Single colonies formed after culturing were identified by Bifidobacterium specific primers for molecular biology. Results After isolation and culture, the detection rate of Bifidobacterium in the plaque and saliva samples of the experimental group was significantly higher than that of the control group (P <0.05). A single colony was selected randomly from the experimental group for Bifidobacterium and sequenced after PCR amplification. The results showed that Bifidobacterium bifidum accounted for 81.67% and Bifidobacterium aldus accounted for 6.67% Bifidobacteria accounted for 11.66%; saliva samples of 60 colonies of Bifidobacterium odoratum accounted for 88.33%, Bifidobacterium dinosaur accounted for 3.34%, bifidobacteria accounted for 8.33%. Conclusion Bifidobacteria have a certain correlation with the occurrence of severe dental caries in children. The main strains of Bifidobacterium oralis in children are Bifidobacterium binatus, Bifidobacterium dentate and Bifidobacterium bifidum, of which the highest proportion of Bifidobacterium bifidum, Is an active dental caries in children.