Dou Qingya, Zou Mingxiang, Li Jun, Wang Haichen, Hu Yongmei, Liu Wen'en
Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha 410008, China.
Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2017 Apr 28;42(4):426-433. doi: 10.11817/j.issn.1672-7347.2017.04.010.
To investigate relationship between AdeABC efflux pump and resistance of Acinetobacter baumannii against carbapenem. Methods: Carbapenem-resistant strains were acquired from multistep selection resistance test by meropenem in vitro. The quantitation test for sensitivities of strains before and after induction was determined by the E-test, and carbonylcyanide-m-chlorophenylhydrazone (CCCP) inhibition test was used to screen efflux pump. PCR, sequencing analysis, or real-time PCR was used to analyze the changes of regulatory genes adeR and adeS of the AdeABC efflux pump system, or expressions of adeA, adeB, adeR, and adeS in the strains before and after induction, respectively. Results: The minimal inhibitory concentrations (MICs) of meropenem were at 0.38 μg/mL and 0.25 μg/mL in parental sensitive strain S25595 and S7257, respectively, and the MICs of meropenem for both S25595 and S7257 after induction were more than 32 μg/mL. Compared with parental sensitive strains, the expression level of adeA, adeB, adeR, and adeS mRNA were elevated from 2.45 to 9.44 times, but there were no gene mutations or insertion sequences in the regulatory gene adeS and adeR. Conclusion: High expression of the AdeABC efflux pump system in Acinetobacter baumannii is closely associated with meropenem resistance. The upregulation of adeA and adeB expression is not due to gene mutations in the regulatory gene adeS and adeR and other mechanisms might account for it.
探讨AdeABC外排泵与鲍曼不动杆菌对碳青霉烯类耐药性之间的关系。方法:通过美罗培南体外多步筛选耐药试验获得碳青霉烯类耐药菌株。采用E-test法测定诱导前后菌株的敏感性定量试验,并用羰基氰化物间氯苯腙(CCCP)抑制试验筛选外排泵。分别采用PCR、测序分析或实时PCR分析AdeABC外排泵系统调控基因adeR和adeS的变化,或诱导前后菌株中adeA、adeB、adeR和adeS的表达。结果:亲本敏感菌株S25595和美罗培南的最低抑菌浓度(MICs)分别为0.38μg/mL和0.25μg/mL,诱导后S25595和S7257的美罗培南MICs均大于32μg/mL。与亲本敏感菌株相比,adeA、adeB、adeR和adeS mRNA的表达水平提高了2.45至9.44倍,但调控基因adeS和adeR中未发现基因突变或插入序列。结论:鲍曼不动杆菌中AdeABC外排泵系统的高表达与美罗培南耐药密切相关。adeA和adeB表达上调并非由于调控基因adeS和adeR的基因突变,可能由其他机制引起。