多重耐药鲍曼不动杆菌外排泵表型和基因型的研究

Relationship between efflux pump phenotype and genotype in multiple drug Acinetobacter baumannii

目的：探讨多重耐药鲍曼不动杆菌外排泵表型和外排泵基因表达情况在耐药方面的作用。方法应用K-B法检测河北医科大学第一医院96株多重耐药鲍曼不动杆菌对15种抗菌药物的耐药情况，应用微量肉汤稀释法通过加入羰基氰氯苯腙（ CCCP）泵抑制剂检测多重耐药鲍曼不动杆菌外排泵表型，并用PCR扩增进行外排泵蛋白基因的检测和测序。结果96株多重耐药鲍曼不动杆菌对喹诺酮类、头孢菌素类、氨基糖苷类、四环素类抗菌药物的耐药率为70．8%-94．8%。其中有34株外排泵表型阳性；外排泵表型阳性的34株鲍曼不动杆菌检测到 adeB、adeR、adeS、adeJ、adeE、adeM 基因分别有33株、32株、33株、33株、0株、33株，检出阳性率分别为97．06%、94．12%、97．06%、97．06%、0、97．06%；对adeB、adeR、adeS基因进行测序，经比对，所测序列与GenBank中序列同源性为100%。结论主动外排泵基因是鲍曼不动杆菌发生多重耐药的重要因素之一。

Objective To expore multi-drug resistant Acinetobacter baumannii efflux pump phenotype and efflux pump gene expression in the resistant isolates. Methods Application of K-B method to detect 96 strains isolated from the First Hospital of Hebei Medical University multi-drug resistant Acinetobacter baumannii′ resistance to 15 kinds of antibacterial drugs, detecting multi-drug resistant Acinetobacter baumannii efflux pump phenotype with broth microdilution method by the addition of carbonyl cyanide chlorobenzene hydrazone （ CCCP） pump inhibitors,using PCR amplification and sequencing to study efflux pump protein gene sequence characteristics . Results The Acinetobacter baumannii resistance rate of 96 strains to quinolones, cephalosporins, aminoglycosides, tetracyclines were 70. 8%-94. 8%.There were 34 positive efflux pump phenotypes in 96 multi-drug resistant Acinetobacter baumannii strains, including 33 adeB strains, 32 adeR strains, 33 adeS strains, 33 adeJ strains,0 adeE strain,33 adeM strains, positive detection rate were 97. 06%, 94. 12%, 97. 06%, 97. 06%, 0, 97. 06%, respectively. By sequence comparison, adeB, adeR and adeS genes sequence homology was 100% in the GenBank. Conclusion Active efflux pump gene perturbation is one of the important factors in multi-drug resistant Acinetobacter baumannii.