中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2008年
2期
162-165
,共4页
菌斑生物被膜%链球菌%寡核苷酸探针%荧光原位杂交
菌斑生物被膜%鏈毬菌%寡覈苷痠探針%熒光原位雜交
균반생물피막%련구균%과핵감산탐침%형광원위잡교
Dental biofilm%Streptpcoccus%Oligonucleotide probe%Fluorescence in situ hybridization(FISH)
目的 检测菌斑生物被膜初期形成过程中的变形链球菌(Streptpcoccus mutans)、远缘链球菌(Streptpcoccus sobrinus)和血链球菌(Streptpcoccus sanguis).方法 从植入釉质磨片表面获得完整的菌斑生物被膜标本,应用激光共聚焦扫描显微镜和荧光原位杂交技术相结合的方法,对天然菌斑生物被膜形成初期的变形链球菌、远缘链球菌和血链球菌进行原位的、实时的动态观察,通过连续断层扫描及三维重建,观察这3种菌在天然菌斑生物被膜中的空间分布,测量3种细菌的扫描厚度.实验中的扫描厚度结果采用方差分析方法进行统计处理,SPSS11.5统计软件辅助完成,α设在0.05.结果 在激光共聚焦扫描显微镜下观察生物被膜呈三维立体结构,形态多样,生物被膜内层和外层细菌较稀疏,中间层较密集,细菌之间可见许多黑色空隙,贯穿整个生物被膜.菌斑生物被膜变形链球菌、远缘链球菌和血链球菌在菌斑生物被膜形成初期的平均扫描厚度随时间延长而增加,1 h时平均扫描厚度分别为20.43、11.50、14.76 μm,到24 h时平均厚度最大,分别为70.25、75.40、79.98 μm.结论 应用荧光原位杂交技术结合激光共聚焦扫描显微镜,可以快速、灵敏地检测出菌斑生物被膜变形链球菌、远缘链球菌和血链球菌.
目的 檢測菌斑生物被膜初期形成過程中的變形鏈毬菌(Streptpcoccus mutans)、遠緣鏈毬菌(Streptpcoccus sobrinus)和血鏈毬菌(Streptpcoccus sanguis).方法 從植入釉質磨片錶麵穫得完整的菌斑生物被膜標本,應用激光共聚焦掃描顯微鏡和熒光原位雜交技術相結閤的方法,對天然菌斑生物被膜形成初期的變形鏈毬菌、遠緣鏈毬菌和血鏈毬菌進行原位的、實時的動態觀察,通過連續斷層掃描及三維重建,觀察這3種菌在天然菌斑生物被膜中的空間分佈,測量3種細菌的掃描厚度.實驗中的掃描厚度結果採用方差分析方法進行統計處理,SPSS11.5統計軟件輔助完成,α設在0.05.結果 在激光共聚焦掃描顯微鏡下觀察生物被膜呈三維立體結構,形態多樣,生物被膜內層和外層細菌較稀疏,中間層較密集,細菌之間可見許多黑色空隙,貫穿整箇生物被膜.菌斑生物被膜變形鏈毬菌、遠緣鏈毬菌和血鏈毬菌在菌斑生物被膜形成初期的平均掃描厚度隨時間延長而增加,1 h時平均掃描厚度分彆為20.43、11.50、14.76 μm,到24 h時平均厚度最大,分彆為70.25、75.40、79.98 μm.結論 應用熒光原位雜交技術結閤激光共聚焦掃描顯微鏡,可以快速、靈敏地檢測齣菌斑生物被膜變形鏈毬菌、遠緣鏈毬菌和血鏈毬菌.
목적 검측균반생물피막초기형성과정중적변형련구균(Streptpcoccus mutans)、원연련구균(Streptpcoccus sobrinus)화혈련구균(Streptpcoccus sanguis).방법 종식입유질마편표면획득완정적균반생물피막표본,응용격광공취초소묘현미경화형광원위잡교기술상결합적방법,대천연균반생물피막형성초기적변형련구균、원연련구균화혈련구균진행원위적、실시적동태관찰,통과련속단층소묘급삼유중건,관찰저3충균재천연균반생물피막중적공간분포,측량3충세균적소묘후도.실험중적소묘후도결과채용방차분석방법진행통계처리,SPSS11.5통계연건보조완성,α설재0.05.결과 재격광공취초소묘현미경하관찰생물피막정삼유입체결구,형태다양,생물피막내층화외층세균교희소,중간층교밀집,세균지간가견허다흑색공극,관천정개생물피막.균반생물피막변형련구균、원연련구균화혈련구균재균반생물피막형성초기적평균소묘후도수시간연장이증가,1 h시평균소묘후도분별위20.43、11.50、14.76 μm,도24 h시평균후도최대,분별위70.25、75.40、79.98 μm.결론 응용형광원위잡교기술결합격광공취초소묘현미경,가이쾌속、령민지검측출균반생물피막변형련구균、원연련구균화혈련구균.
Objective To examine Streptpcoccus mutans,Streptpcoccus sobrinus and Streptpcoccus sanguis in the early formation of native dental plaque biofilm. Methods An experimental dental plaque biofilm model in the oral cavity was established using enamel slabs. The spatial distribution of S. mutans, S.sobrinus and S. sanguis in the early colonization of dental plaque biofilms on the enamel surface was observed bv in situ, real-time and dynamic observations and optical sections utilizing confocal laser scanning microscopy(CLSM) and fluorescence in situ hybridization(FISH). The experiment data were analyzed with One-Way AVOVA, α=0.05 using SPSS11.5. Results Dental biofilm had a certain degree of thickness and various forms in three-dimensioned structure. The bacteria in the structure were sparse at the inner layers and the outer layers. In the middle layers the bacteria were closely compacted. There were many voids traversing from the outside of the biofilm to the enamel surface. At the initial stage of dental biofilm formation, the scanned average thickness of S. mutans,S. sobrinus and S. sanguis increased with time elapsing,the mean thicknesses of 1 h biofilms were 20.43 μm,11.50 μm and 14.76 μm,respectively,and those of 24 h were the thickest in terms of average level,the mean values were 70.25 μm,75.40 μm and 79.98 μm,respectively. Conclusion The fluorescence in situ hybridization combined with CLSM are thought to be convenient and sensitive to detect S. mutans, S. sobrinus and S. sanguis in the dental plaque biofilms.
