昆明地区耐碳青霉烯铜绿假单胞菌10种膜蛋白编码基因表达的实验研究  

Experimental Study on Expression of Carbapenem Resistant Pseudomonas Aeruginosa’s 10 Membrane Protein Coding Genes in Kunming

作　　者：卢赞[1] 赵红燕[1] 李春付 尹利民 任宝军[1] 宋贵波[2] 杨旭[3] LU Zan;ZHAO Hongyan;LI Chunfu;YIN Limin;REN Baojun;SONG Guibo;YANG Xu(the First People’s Hospital of Kunming,Kunming 650221,China;the First Affiliated Hospital of Kunming Medical University,Kunming 650032,China;the Second Affiliated Hospital of Kunming Medical University,Kunming 650101,China)

机构地区：[1]昆明市第一人民医院,昆明650221 [2]昆明医科大学第一附属医院,昆明650032 [3]昆明医科大学第二附属医院,昆明650101

出　　处：《现代检验医学杂志》2025年第1期7-12,共6页Journal of Modern Laboratory Medicine

基　　金：昆明市卫生健康委员会卫生科研项目(2022-11-01-022)。

摘　　要：目的 掌握昆明地区碳青霉烯耐药铜绿假单胞菌(CRPA)膜蛋白表达的分子流行情况,为临床合理用药及外排泵抑制剂的应用提供依据。方法 收集昆明地区四所医院2022年10月~2023年8月分离的铜绿假单胞菌,使用SYBR-PCR法定量检测10种膜蛋白编码基因mRNA相对表达量(RE),包括mexA,B,C,D,E,F,X,Y及oprD,M。根据头孢他啶(CAZ)、头孢吡肟(CFP)、亚胺培南(IPM)、美罗培南(MEM)耐药表型组合将菌株分为5组,包括全敏感组(Ⅰ组),全耐药组(Ⅱ组),IPM,MEM耐药、CAZ及CFP敏感组(Ⅲ组),IPM耐药、MEM非耐药(敏感或中介)组(Ⅳ组)和IPM,MEM耐药、CAZ和CFP非耐药组(Ⅴ组),分析不同耐药表型组间各膜蛋白编码基因的RE中位数情况。结果 共收集108株铜绿假单胞菌,Ⅰ组24株作为对照,84株为碳青霉烯耐药组,包括Ⅱ组32株,Ⅲ组22株,Ⅳ组13株和Ⅴ组17株。耐药组mexD,mexE,mexF,mexX和mexY表达高于对照组,差异具有统计学意义(U=409.5~661.0,均P<0.05);mexA,mexB,mexC,oprD,oprM与对照组比较,差异无统计学意义(U=767.0~1 004.5,均P>0.05)。各膜蛋白编码基因RE在不同医院来源菌株间的表达,差异无统计学意义(H=0.914~7.407,均P>0.05)。四组不同表型中,mexA和oprM RE在各组无规律分布与对照组比较差异无统计学意义(U_(mexA)=95.0~264.0,U_(oprM)=143.0~331.0);各组mexC RE均低于对照组,但差异无统计学意义(U=134.0~344.5,均P>0.05);mexE和mexY RE均高于对照组(U_(mexE)=48.0~230.0,U_(maxY)=83.0~184.0),mexB在Ⅳ组中低于对照组(U=72.0),差异具有统计学意义(均P<0.05)。mexD和mexF表现一致,在Ⅲ,Ⅳ,Ⅴ组表达高于对照组(U_(mexD)=34.0~102.0,U_(mexF)=65.0~113.0),mexX在Ⅱ,Ⅳ,Ⅴ组表达高于对照组(U=164.0,58.0,111.0),oprD仅在Ⅲ组表达低于对照组(U=140.0),差异具有统计学意义(均P<0.05);oprD在Ⅱ,Ⅳ,Ⅴ组中表达虽低于对照组,但差异无统计学意义(U=381.0,102.0,144.0,均P>0.05)。结论 mexCD,mexEF,mexXY是昆明地区CRPA主要外排泵Objective To understand the membrane protein molecular epidemiology of carbapenem-resistant Pseudomonas aeruginosa(CRPA)in the region,and provide some evidence for rational drug use or application of efflux pump inhibitors.Methods Collected Pseudomonas aeruginosa isolated from four hospitals in the region from October 2022 to August 2023,and used SYBR-PCR method to quantitatively detect the relative mRNA expression(RE)levels of 10 membrane protein coding genes,including mexA,B,C,D,E,F,X,Y,and oprD,M.Then categorized the strains into five groups based on ceftazidime,cefepime,imipenem,and meropenem resistance phenotype combination,including the compassionate group(Group Ⅰ),Group Ⅱ with full resistance,IPM,MEM resistant,CAZ and CFP sensitive groups(Group Ⅲ),IPM resistance,MEM non-resistance(sensitive or intermediate)group(Group Ⅳ),IPM,MEM resistance,CAZ and CFP non-resistance groups(Group Ⅴ).The median RE of each membrane protein-coding gene was analyzed.Results A total of 108 strains of Pseudomonas aeruginosa were collected,with 24 strains in Group Ⅰ as controls and 84 strains in the carbapenem resistant group,including 32 strains in Group Ⅱ,22 strains in Group Ⅲ,13 strains in Group Ⅳ,and 17 strains in Group Ⅴ.The expression of mexD,mexE,mexF,mexX and mexY in the drug-resistant group was higher than that in the control group,and the differences were statistically significant(U=409.5~661.0,all P<0.05).There was no statistically significant difference in mexA,mexB,mexC,oprD and oprM with the control group(U=767.0~1004.5,all P>0.05).There was no significant difference in the expression of RE genes encoding various membrane proteins among strains from different hospitals(H=0.914~7.407,all P>0.05).Among the four different phenotypes,there was no statistically significant difference in the irregular distribution of mexA and oprM RE between each group and the control group(U_(mexA)=95.0~264.0,U_(oprM)=143.0~331.0).The mexC RE in each group was lower than that in the control group,but the differences we

关 键 词：铜绿假单胞菌 碳青霉烯耐药 外排泵 膜蛋白 

分 类 号：R378.991[医药卫生—病原生物学] Q786[医药卫生—基础医学]