口腔常见微生物不同定量分析方法的比较  

Comparative study on different methods for quantitative analysis of oral common microorganisms

作　　者：张明珠[1] 李超伦[2] 姜云涛[1] 姜葳[1] 梁景平[1] 叶冬霞[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152-155,共4页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(30572039)~~

摘　　要：目的探索口腔微生物定量分析的理想方法。方法选择3种口腔常见微生物变异链球菌、牙龈卟啉单胞菌和具核梭杆菌的标准菌种厌氧培养24 h,经倍比稀释,涂板菌落形成单位(CFU)计数。将相应体积的菌液固定后,分别运用3种非培养方法对上述细菌进行定量分析(实时PCR、激光共聚焦显微镜和流式细胞仪)。结果细菌定量结果由大至小排列,分别为实时PCR分析、激光共聚焦显微镜观察、CFU计数、流式细胞仪检测。每种细菌的实时PCR标准曲线均显示具备较好的线性关系。结论本研究中的3种非培养方法均适用于口腔微生物的定量分析。其中实时PCR分析和激光共聚焦显微镜观察的结果显著优于传统的CFU计数方法,而以实时PCR的敏感度最高。Objective To determine an ideal method for quantitative analysis of oral microorganisms. Methods Three oral common microorganisms, Streptococcus mutans（ S. mutans ） , Porphyromonas gingivalis（ P. gingivalis ） and Fusobacterium nucleatum （ F. nucleatum） were selected and cultured in vitro for 24 h. The total number of colony-forming units （CFU） was determined on the basis of serial dilution on selective media. The individual bacterial species were quantitatively analyzed by three non-culture methods （real-time PCR, laser scanning confocal microscopy and flow cytometry）. Results The quantitative analysis indicated the following order in the total number of bacterial species： real-time PCR 〉 laser scanning confocal microscope 〉 CFU count 〉 flow cytometry. A strong linear relationship was observed in the standard curve of real-time PCR of each bacteria. Conclusion These three non-culture methods can be used in the quantitative analysis of oral microorganisms. Real-time PCR and laser scanning confocal microscopy are better than the traditional culture-based CFU count, and real-time PCR is the most sensitive method.

关 键 词：实时PCR 激光共聚焦显微镜 流式细胞仪 菌落形成单位 

分 类 号：R780.2[医药卫生—口腔医学]