某院4株携带Ⅰ类整合酶基因的铜绿假单胞菌对碳青霉烯类抗菌药物的耐药机制研究  

Study on the resistance mechanism of 4 strains of Pseudomonas aeruginosa with classⅠintegras gene to carbapenem antibacterials in the hospital

作　　者：何蕾 阴晴[1] 周亚玲 吴瑶 HE Lei;YIN Qing;ZHOU Yaling;WU Yao(Department of Clinical Laboratory,Affiliated Hospital of Jiangsu University,Zhenjiang,Jiangsu 212001,China)

机构地区：[1]江苏大学附属医院检验科,江苏镇江212001

出　　处：《国际检验医学杂志》2020年第23期2871-2875,2881,共6页International Journal of Laboratory Medicine

基　　金：江苏省镇江市2017年度科技项目(SH2017024);江苏省2018年度预防医学科研课题(Y2018108)。

摘　　要：目的分析该院4株不同耐药程度的Ⅰ类整合酶基因(IntⅠ)阳性铜绿假单胞菌(PA)的相关耐药机制,从而进一步研究不同耐药机制之间的相互作用。方法采用qPCR检测头孢他啶(CAZ)、亚胺培南(IMP)诱导下的4株PA的IntⅠmRNA的相对表达水平;采用PCR检测4株PA的β-内酰胺酶(blaTEM、blaNDM-1、blaVIM、blaIMP)、主动外排系统(MexAB-OprM)、外膜蛋白(OprD2)的表达水平,并采用qPCR检测4株PA的OprM、OprD2的表达水平;采用1%结晶紫染色检测4株PA的生物膜形成能力。结果编号为PA01、PA02、PA04的菌株在CAZ、IMP诱导下,IntⅠmRNA的相对表达水平均较对照组高(P<0.05),PA01、PA02的IntⅠmRNA的相对表达水平高于PA04,组间两两比较,差异均有统计学意义(P<0.05),并且在一定浓度范围内,IntⅠmRNA的相对表达水平随着抗菌药物的浓度增加而增加。但PA03菌株的IntⅠmRNA的相对表达水平较PA01、PA02、PA04低,组间比较差异无统计学意义(P>0.05)。PA01、PA02检测到blaTEM,而blaNDM-1、blaVIM和blaIMP均未被检测到;PA01、PA02的外排泵相对表达水平高于PA03、PA04(P<0.05);PA01、PA02的OprD2 mRNA的相对表达水平低于PA03、PA04(P<0.05);PA01、PA02的生物膜形成能力弱于PA03、PA04,且PA03菌株的生物膜形成能力最强。结论PA01、PA02、PA04的IntⅠ在抗菌药物的选择性压力下表达增强,并且PA01、PA02对碳青霉烯类耐药主要由IntⅠ高表达、外排泵过表达及膜通透性降低介导,与β-内酰胺酶及生物膜形成关系不大。Objective To Analyze the related resistance mechanism of 4 strains of different resistance levels of classⅠintegrase gene(IntⅠ)positive Pseudomonas aeruginosa(PA),so as to further study the interaction between different resistance mechanisms.Methods The relative expression levels of IntⅠmRNA in 4 PA strains induced by ceftazidime(CAZ)and imipenem(IMP)were detected by qPCR;β-lactamases(blaTEM,blaNDM-1,blaVIM,blaIMP),the expression levels of active efflux system(MexAB-OprM)and outer membrane protein(OprD2)were detected by qPCR,the expression levels of OprM and OprD2 in 4 PA strains were detected by qPCR;The biofilm formation ability of 4 PA strains was detected by 1%crystal violet staining.Results Under the induction of CAZ and IMP,the relative expression levels of IntⅠmRNA in PA01,PA02 and PA04 was higher than that in the control group(P<0.05),and that of PA01 and PA02 was higher than that in PA04.There was a significant difference between the two groups(P<0.05).In a certain concentration range,the relative expression level of IntⅠmRNA increased with the increase of antibiotic concentration.However,the relative expression level of IntⅠmRNA in PA03 was lower than that in PA01,PA02 and PA04,and there was no significant difference between the groups(P>0.05).The blaTEM gene was detected in PA01 and PA02,but blaNDM-1,blaVIM and blaIMP were not detected in all strains.The relative expression level of efflux pump in PA01 and PA02 was higher than that in PA03 and PA04(P<0.05).The relative expression level of OprD2 in PA01 and PA02 was lower than that in PA03 and PA04(P<0.05).The biofilm forming ability of PA01 and PA02 was weaker than that of PA03 and PA04,and the biofilm forming ability of PA03 was the strongest.Conclusion The expression of IntⅠin PA01,PA02 and PA04 is increased under the selective pressure of antibiotics,and the resistance of PA01 and PA02 to carbapenems is mainly mediated by the high expression of IntⅠ,the overexpression of efflux pump and the decrease of membrane permeability,which is

关 键 词：铜绿假单胞菌 Ⅰ类整合酶基因 β-内酰胺酶 主动外排泵 外膜蛋白 

分 类 号：R446.5[医药卫生—诊断学]