[gbk] 鲍曼不动杆菌检测分析手册[1](4)

纸片扩散法使用MH琼脂培养基，接种前菌量调至0.5麦氏标准浓度，培养条件为35℃±2℃，在大气环境中培养20～24小时。具体操作步骤请参考CLSI/NCCLS M2-A8 标准 “Performance Standards for Antimicrobial Disk Susceptibility Tests”。

（2）E-Test法

E-Test法与NCCLS的稀释法相比其操作较为简便，MIC值更为精确。瑞典AB BlODISK公司生产的E-Test试纸条已有很多种抗生素可供选择，所用的培养基和培养条件与琼脂稀释法相同，其结果判读同稀释法MIC标准。

原理：载体塑料条5×50 mm，一面标有ug/ml为单位的MIC读数，另一面则载有干燥而稳定呈连续指数梯度的抗生素，浓度一般为0.016-256 ug/ml或0.002-32 ug/ml。 当条贴到接种好的琼脂平板表面，条上抗生素立即释放出来，条下方形成连续梯度的浓度。过夜孵育的细菌沿载体条为中心呈椭圆形抑菌圈，椭圆形抑菌圈与载体条交界处即可读出MIC值（ug/ml）。

4． 结果判读和解释

结果判读参照NCCLS标准. 5． 耐药基因检测

不动杆菌耐药机制复杂，主要包括以下几个方面：

（1）产生灭活酶，目前已经发现的与不动杆菌耐药性相关的灭活酶主要包括：苯唑西林酶（oxacillinases，OXA）、金属β—内酰胺酶(metallo-β

-lactamase, MBL)、头孢菌素酶(AmpC)酶和超广谱β—内酰胺酶(extended-spectrumβ-lactamase, ESBLs)。

（2）细菌外膜通透性的改变，

（3）抗生素作用靶位的改变。

（4）细菌细胞膜对抗生素的主动外排机制。

其中灭活酶的产生和和产酶基因的传递又与整合子的作用密切相关。

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