应用新一代高通量测序技术探讨含氟涂料对患龋乳前牙菌斑微生物的影响  

Study on effects of fluoride varnish on plaque biofilm microbial diversity of anterior deciduous teeth with dental caries using sequencing analysis

作　　者：杨琳[1] 张璐 李延平[1] 

机构地区：[1]上海市杨浦区中心医院,上海200090

出　　处：《口腔材料器械杂志》2017年第4期178-183,共6页Chinese Journal of Dental Materials and Devices

摘　　要：目的应用新一代高通量测序技术,了解含氟涂料处理患龋乳前牙前后菌斑微生物的变化。方法选取20名乳前牙患龋的学龄前儿童,菌斑生物膜样本采自含氟涂料处理前和处理后3天、7天和30天的乳前牙患龋牙面;抽提全基因组DNA后,用16S r RNA基因V4-V5高变区为引物扩增后,运用Mi Seq测序平台测序并分析。结果 4个时间点菌斑生物膜微生物多样性Shannon指数的比较表明,涂氟后3天、后7天比涂氟前的Shannon指数更低(P=0.011;P=0.016),提示菌斑生物膜的微生物多样性降低;涂氟后30天和涂氟前的Shannon指数持平(P>0.05),提示涂氟后菌斑生物膜的微生物多样性恢复至涂氟前的状态;PCA分析也进一步表明和Shannon指数一致;4个时间点优势菌属的比较表明涂氟后3天、后7天、后30天和涂氟前优势菌属的种类和相对丰度均没有差异,说明含氟涂料对菌斑生物膜的多样性的抑制作用没有特异性。结论含氟涂料对患龋乳前牙牙面菌斑微生物在一定时间内有明显的抑制作用,抑制作用无特异性,微生物多样性经过一段时间还能恢复至原有状态。Objective To investigate the changes of plaque biofilm microbial diversity before and after fluoride varnish treatment in anterior deciduous teeth with caries using sequencing analysis. Methods 20 preschool children with caries on anterior deciduous teeth were included in this study. Supragingival plaque biofihn samples were collected before fluoride varnish treatment, and at 3 days, 7 days and 30 days after fluoride varnish treatment. Plaque biofilm DNA was extracted, used to generate DNA amplicons of the V4-V5 hypervariable region of the bacterial 16S rRNA gene, and subjected to MiSeq sequencing. Results Sequencing depth was reasonable. The Shannon microbial diversity index showed statistically significant difference both 3 days and 7 days after treatment vs. before treatment （P=O.O 11, P=O.O 16）. The Shannon microbial diversity index showed no statistically signifcant difference before vs. 30 days post treatment （P〉0.05）. The results revealed less microbial diversity 3 days and 7 days post treatment, and microbial diversity returned to the original level 30 days after fluoride varnish treatment. Similar results were found by Principal Component Analysis （PCA）. There was no difference in the species and relative abundance of the predominant bacteria by genus at 3 days, 7 days, 30 days after fluoride varnish treatment,and the results showed that the inhibitory effectof fluoride varnish was not specific. Conclusion Fluoride varnish has non-specific inhibitory effects on plaque biofilm microbial diversity in anterior deciduous teeth with caries during a short period of time, and plaque biofilm microbial diversity was able to return to the original level after 1 month.

关 键 词：含氟涂料 龋齿 菌斑生物膜 微生物多样性 二代高通量测序技术 

分 类 号：R788.1[医药卫生—口腔医学]