原子力显微镜观察耐药铜绿假单胞菌株药物作用前后的细菌表面特征变化  

Surface characteristics of drug-resistant Pseudomonas aeruginosa strain under atomic force microscope before and after treatment with drugs

作　　者：李忠新[1] 张春雷[1] 周小梅[1] 蓝志明[1] 李瑜珍[1] 张书楠[1] 陈萍[1] 吕琪[1] 黄志伟[1] 

机构地区：[1]深圳市中医院检验科,广东深圳518001

出　　处：《第三军医大学学报》2010年第15期1645-1648,共4页Journal of Third Military Medical University

摘　　要：目的从临床分离1株耐β-内酰胺类铜绿假单胞菌(Pseudomonas aeruginosa,PA)菌株,观察其体外耐β-内酰胺类抗菌药物后发生的细菌表面特征变化。方法从伤口分泌物中分离铜绿假单胞菌并采用K-B法进行药物敏感性实验,筛选出1株耐β-内酰胺类抗菌药物的菌株;通过革兰染色和原子力显微镜(atomic force microscope,AFM)观察K-B实验前后细菌形态和表面特征的变化。结果筛选出了1株耐β-内酰胺类抗菌药物的菌株,通过革兰染色和原子力显微镜观察到K-B实验后细菌菌体均变长,能达到K-B实验前的10倍,宽度略微瘦小,即长5.0～25.0μm,宽0.3～0.8μm,表现出L型的细菌形态;革兰阴性的染色特性未发生改变;在500 nm×500 nm的扫面面积下AFM观察到K-B实验后铜绿假单胞菌的表面孔洞增多,显示出粗糙的特征,通过地貌图(K-B实验前深度不大于10 nm,K-B实验后深度大于30 nm)和粗糙度参数分析,结果进一步证实该现象。结论 AFM能观察到K-B实验后铜绿假单胞菌的表面变得更加粗糙的特征,而革兰染色不能观察到这一改变;AFM扫描观察技术可用于细菌表面微观变化观察。Objective To observe the surface characteristics of in vitro β-1actam resistant Pseudomonas aeruginosa after treated with β-1actam resistant antibiotics. Methods A Pseuclomonas aeruginosa strain was isolated from wound secretions with the K-B method to test its susceptibility to β-lactam resistant antibiotics. Morphology of the stain and its surface characteristics were observed with Gram staining under atomic force microscope （AFM）. Results A β-lactam antibiotics resistant Pseudomonas aeruginosa strain was isolated. The strain after Gram staining was 10-fold longer （5.0 to 25.0μm） and slightly less wider （0.3 to 0.8 μm） under atomic force microscope after K-B experiments. The strain was L-shaped. No change was found in the strain after Gram-negative staining. Atomic force microscopy showed more holes on surface of the strain under a scanning area of 500 nm ×500 nm, which were further confirmed on the topographic maps （the depth was less than 10 nm before K-B test and greater than 30 nm after K-B test）. Conclusion Atomic force microscopy can show the rough surface characteristics of Pseudomonas aeruginosa after K-B experiment, but cannot show them after Gram staining, indicating that AFM scanning can be used to observe microchanges on the surface of the strain.

关 键 词：铜绿假单胞菌 Β-内酰胺 原子力显微镜 

分 类 号：R378.991[医药卫生—病原生物学] R446.5[医药卫生—基础医学]