国际口腔医学杂志 ›› 2017, Vol. 44 ›› Issue (4): 411-420.doi: 10.7518/gjkq.2017.04.009
陆笑1, 翁春辉1, 王劲茗2, 刘少娟1, 刘芹1, 林珊1
Lu Xiao1, Weng Chunhui1, Wang Jinming2, Liu Shaojuan1, Liu Qin1, Lin Shan1
摘要: 目的 了解luxS基因敲除,自体诱导物-2(AI-2)密度感应信号缺失对缓症链球菌生物膜致病力表型的影响。方法 常规细菌培养、药敏试验、形态学与生化反应鉴定,16S 核糖体DNA(rDNA)序列同源性分析,获得耐药性缓症链球菌临床分离株。同源重组法敲除luxS基因,构建突变株。哈氏弧菌BB170菌株生物发光实验检测AI-2信号;定量分析临床分离株与突变株生物膜形成量与抗菌药敏感性的差异;荧光黄染液、18909荧光增白染液、L7012死/活菌染液染色,激光共聚焦显微镜扫描,了解生物膜结构、胞外多糖、死/活菌分布差异;临床分离株上清液、4,5-二羟基-2,3-戊二酮(DPD)补偿生长实验确认突变株表型改变的原因。实验数据行Kolmogorov-Smirno非参数检验。结果 luxS基因突变株与临床分离株生物膜形成量存在显著性差异(P<0.001);氨苄西林、环丙沙星、四环素干预条件下,突变株生物膜形成量显著降低。上清液、DPD补偿后,突变株生物膜形成能力和结构可以恢复。突变株生物膜结构变疏松,对氨苄西林、环丙沙星、四环素的敏感性增加。结论 luxS/AI-2密度感应信号缺失,导致耐药性缓症链球菌生物膜形成量下降,抗菌药敏感性增加,致病力表型发生改变。
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