细菌生物膜检测方法改进与应用  

Improved method of bacterial biofilm detection and its application

作　　者：尤理想 赵青[1] 周敏[1] 汪意涛 吴淑燕[1] 李嫄渊[1] 黄瑞[1] 

机构地区：[1]苏州大学病原生物学系,江苏苏州215000

出　　处：《实验技术与管理》2015年第3期72-76,共5页Experimental Technology and Management

基　　金：江苏省自然科学基金项目(No.2011286);江苏省高等学校创新性实验计划项目(No.2012yb017)

摘　　要：针对结晶紫染色法为细菌生物膜(biofilm,BF)检测的经典方法存在的不足,通过增加乙酸用量和减少洗菌次数等进行改进后,BF测定更为准确。另尝试采用多模式小动物活体成像系统检测发光菌的BF形成,并利用培养板和小圆玻片探究BF在不同载体上的形成情况,以优化小动物活体成像系统检测方法。用改进后的2种方法摸索鼠伤寒沙门菌BF形成的最适浓度和时间,结果发现,接种浓度为107 CFU/mL、培养72h时检测效果最佳。在BF检测方法优化的基础上。探究鼠伤寒沙门菌质粒对BF形成的影响,结果发现质粒可促进BF的形成。不同亚抑菌浓度双黄连对鼠伤寒沙门菌BF作用的实验结果显示,1/4 MIC(MIC为最小抑菌浓度)双黄连对BF形成的抑制作用显著低于1/2 MIC和1/8 MIC,表明双黄连的抑菌作用与浓度不成正比关系,需严格控制临床用药剂量。Crystal violet staining is a classical method for the detection of bacterial biofilm（BF）,but remains deficiencies in mechanical process.In this study,the method is improved by increasing the amount of acetic acid and reducing washing times.The multi-mode small animal imaging system is used to detect the formation of luminous bacteria,using small circular glass and plastic plates to explore the formation of BF on different carriers to optimize the detection methods.By exploring the optimal concentration and culture time of Salmonella BF formation using the inmentioned improved detection methods,it is found that BF detected effect is the best when inoculating 107 CFU/ml of bacteria and cultured for 72 h.On the basis of the BF detection method optimization,this article explores the effects of Salmonella plasmid on biofilm formation,and the results show that the plasmid has a role in promoting BF formation.The experimental results of different subMIC of Shuanghuanglian on Salmonella BF formation show that 1/4MIC of Shuanghuanglian has the minimum inhibition effect to Salmonella BF formation compared with 1/2MIC and 1/8MIC of Shuanghuanglian,which hints at that the inhibition effect of Shuanghuanglian is not in direct proportion to its concentration and the dosage of clinical medicine needs to be strictly controlled.

关 键 词：细菌生物膜 结晶紫染色 多模式小动物活体成像系统 鼠伤寒沙门菌质粒 双黄连 

分 类 号：Q939.93[生物学—微生物学]