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目的观察外排泵抑制剂对多重耐药不动杆菌耐药水平的降低作用,并扩增外排泵蛋白编码基因,以探讨不动杆菌多重耐药与细胞膜主动外排作用的关系。方法用琼脂稀释法测定30株鲍曼不动杆菌对环丙沙星、四环素、阿米卡星和亚胺培南的最小抑制浓度(MIC)值,并观察在含10μg/ml利血平或25μg/ml羰基氰氯苯腙(CCCP)条件下MIC值的变化。用PCR法扩增外排泵编码基因adeB。结果以环丙沙星、四环素、阿米卡星和亚胺培南作为底物,分别有12、5、6、6株菌在10μg/mlCCCP或含25μg/ml利血平的条件下MIC值降低4倍或4倍以上。6、11、17号菌都同时对3种药物有明显的外排作用。以利血平和CCCP作为抑制剂所得的结果一致。18株菌经PCR扩增得到与目的片段大小一致的阳性片段,经测序证实与GenBank登录号为AF370885的鲍曼不动杆菌RND族药物外排泵编码基因adeB的同源性达99%。该序列已在GenBank登录,登录号为DQ294294。结论主动外排作用是鲍曼不动杆菌多重耐药的重要原因之一。
Objective To observe the effect of efflux pump inhibitors on the drug resistance of multidrug-resistant Acinetobacter (Mycobacterium tuberculosis) and to amplify the coding genes of efflux pump to explore the relationship between the multi-drug resistance of Acinetobacter and active efflux of cell membrane. Methods The minimal inhibitory concentrations (MICs) of 30 strains of Acinetobacter baumannii on ciprofloxacin, tetracycline, amikacin and imipenem were determined by agar dilution method. The inhibitory concentrations of 10 μg / ml reserpine or MIC of 25μg / ml cyanocarbonyl chloride (CCCP). Amplification of efflux pump encoding gene adeB by PCR method. RESULTS: Ciprofloxacin, tetracycline, amikacin and imipenem were used as substrates respectively. MIC values of 12, 5, 6, and 6 strains at 10 μg / ml CCCP or 25 μg / ml reserpine Reduce 4 times or more. 6,11,17 bacteria have obvious efflux of the three drugs at the same time. The results obtained with reserpine and CCCP as inhibitors were consistent. 18 strains were amplified by PCR and the positive fragments of the same size as the target fragment were identified. The sequence identity with the gene encoding adeB from the RING gene of the Acinetobacter baumannii GenBank accession number AF370885 was 99%. This sequence has been deposited in GenBank under the accession number DQ294294. Conclusion Active efflux is one of the important reasons for multi-drug resistance of Acinetobacter baumannii.