免疫荧光法检测恒磨窝沟菌斑内变形链球菌

Detection of the mutans Streptococci in the fissure plaque of permament molars

目的检测有龋窝沟与无龋窝沟中变形链球菌的数量,初步探讨窝沟菌斑内变形链球菌数量与窝沟龋损的相互关系。方法首先用变形链球菌制备兔抗变形链球菌的抗血清。窝沟菌斑与兔抗变形链球菌血清共同孵育后与异硫氰酸荧光素标记的羊抗兔IgG进行孵育,再行激光共聚焦显微镜扫描,获取窝沟菌斑中变形链球菌的荧光图像并分析。将牙片进行显微放射照相分析,确定样本有无龋坏,并以此结果将所有样本分为有龋组(10例)和无龋组(14例),进行有龋组和无龋组平均荧光强度和荧光图像的对比分析。结果荧光强度分析显示,有龋组和无龋组平均荧光强度无显著性差异(P>0.05)。不同形态的窝沟,变形链球菌的平均荧光强度不同,I型窝沟较弱,I型窝沟与龋损对应位置与其它位置差别不大;U或V型窝沟内变形链球菌的荧光较强。在有龋的样本中,与龋损对应位置变形链球菌的荧光强度似乎较其它位置强。结论变形链球菌存在于所有样本的窝沟内,其数量可能与窝沟的形态有关,而与窝沟是否有龋无关。

Objective To detect the mutans Streptococci in fissure plaque by confocal laser scanning microscopy （CLSM）. Methods Anti -mutans Streptococcus serum was prepared. Thin fissure plaque specimens were made by sectioning the fissure of fresh extracted molars. The fissure plaques were incubated with anti-serum and fluoreseeinisothioeyanate-labeled goat-anti-rabbit IgG, and then scanned by CLSM . Fissure sections were analyzed by microradiograph. According to the microradiograph results, all specimens were divided into two groups： carious group and non-caries group. The average fluorescence and photograph of mutans Streptococcus was compared between caries and caries-free group. Results No significant difference was noted in the average fluorescence between carious and non- mcaries group. Fluorescence photograph of mutans Streptococcus in the fissures were related to the fissure types. The fluorescence of mutans Streptococcus was weaker in type ＂Ⅰ＂ fissures than in type ＂U＂ and ＂V＂ fissures. Conclusion Mutans Streptococcus was detected in all fissure plaques samples and their levels may be more associated with fissure types than with fissure caries.