实时PCR技术在龋易感儿童牙菌斑致龋菌定量检测中的应用  

Quantitative analysis of cariogenic strains in dental plaque of caries-susceptibile children by real-time PCR

作　　者：梁景平[1] 姜云涛[1] 李超伦[2] 姜葳[1] 

机构地区：[1]上海交通大学医学院第九人民医院牙体牙髓科上海市口腔医学重点实验室,上海200011 [2]上海交通大学医学院第九人民医院牙周科上海市口腔医学重点实验室,上海200011

出　　处：《上海交通大学学报（医学版）》2007年第2期128-132,共5页Journal of Shanghai Jiao tong University:Medical Science

基　　金："十五"国家科技攻关计划(2004BA720A24);国家自然科学基金(30572039)~~

摘　　要：目的 建立定量检测致龋菌的快速、敏感、特异的方法。分析牙菌斑中主要致龋菌数量与乳牙龋失补牙面（dmfs）指数的关系。方法 牙菌斑组织取自45例学龄前儿童，其中无龋（drnfs=0）、中龋（dmfs=4—6）和高龋（dmfs〉8）组各15例。应用SYBR Green I模式的实时PCR技术，对各组儿童牙菌斑中4种主要致龋菌（变异链球菌、远缘链球菌、黏性放线菌和内氏放线菌）进行定量检测，并计算各致龋菌所占总菌的比例。对各组所获数据进行统计学分析。结果 高龋组中4种致龋菌所占总菌的比例均高于无龋组（P〈0．01），其中变异链球菌和远缘链球菌的数量之和所占总菌的比例与dmfs指数具有良好的线性关系。结论 实时PCR技术可作为一种细菌定量检测方法，应用于致龋菌的定量分析和龋齿活跃件的评估。Objective To establish and develop a quick,special and sensitive method for quantifying carigenic strains and assessing the relationship between the level of four-species cariogenic pathogens in dental plaque and dmfs index. Methods Dental plaques were obtained from 45 Chinese pre-school children, 15 in caries-free group （dmfs = 0）, 15 in moderate-caries group （ dmfs=4 -6） and 15 in caries-susceptive group （ dmfs 〉 8）. The amount of the targeted microorganisms（ Streptococcus mutans, Streptococcus sobrinus ,Actinomyces viscosus and Actinomyces naeslundii） and the total number of bacterial cells were determined by real-time PCR based on SYBR-Green I fluorescence. Results The percentages of the four targeted bacteria in high-caries group were significantly higher than those in cariesfree group （ P 〈 0.01 ） , and there was good linear correlation between dmfs index and the sum of the percentages of Streptococcus mutans and Streptococcus sobrinus. Conclusion Real-time PCR can be employed in the identification and enumeration of carigenic strains, which are essential for the accurate evaluation of caries susceptibility and risk of caries development.

关 键 词：实时PCR 龋病易感性 致龋菌 牙菌斑 

分 类 号：R788.1[医药卫生—口腔医学]