[1]卢 赞,赵红燕,李春付,等.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[J].现代检验医学杂志,2025,40(01):7-12.[doi:10.3969/j.issn.1671-7414.2025.01.002]
　LU Zan,ZHAO Hongyan,LI Chunfu,et al.Experimental Study on Expression of Carbapenem Resistant Pseudomonas Aeruginosa’s 10 Membrane Protein Coding Genes in Kunming[J].Journal of Modern Laboratory Medicine,2025,40(01):7-12.[doi:10.3969/j.issn.1671-7414.2025.01.002]
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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()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第40卷
期数:
2025年01期
页码:
7-12
栏目:
论著
出版日期:
2025-01-15

文章信息/Info

Title:
Experimental Study on Expression of Carbapenem Resistant Pseudomonas Aeruginosa’s 10 Membrane Protein Coding Genes in Kunming
文章编号:
1671-7414（2025）01-007-06
作者:
卢 赞1; 赵红燕1; 李春付1; 尹利民1; 任宝军1; 宋贵波2; 杨 旭3
（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）
Author(s):
LU Zan1; ZHAO Hongyan1; LI Chunfu1; YIN Limin1; REN Baojun1; SONG Guibo2; YANG Xu3
（1．the First People’s Hospital of Kunming，Kunming 650221，China；2．the First Affiliated Hospital of Kunming Medical University, Kunming 650032，China；3．the Second Affiliated Hospital of Kunming Medical University， Kunming 650101，China）
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；Q786
DOI:
10.3969/j.issn.1671-7414.2025.01.002
文献标志码:
A
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（UmexA=95.0 ～ 264.0，UoprM=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（UmexE=48.0 ～ 230.0，UmaxY=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（UmexD=34.0 ～ 102.0，UmexF=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 主要外排泵的膜蛋白组合，通过mexD，E，F，X，Y 膜蛋白表达上调加强外排，mexAB-oprM 外排泵与该地区CRPA 碳青霉烯耐药相关性低，oprD 低表达在不产β- 内酰胺酶的菌株中与外排机制共同发挥作用，但在产酶菌株中则未见显著的低表达差异。
Abstract:
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 (UmexA=95.0 ～ 264.0，UoprM =143.0 ～ 331.0). The mexC RE in each group was lower than that in the control group , but the differences were not statistically significant (U=134.0 ～ 344.5, all P>0.05). MeixE and meixY RE wer e both higher than the con trol group, and the d i ff e r ences wer e s tat i s t ical l y s igni f ican t (UmexE =48.0 ～ 230.0,UmexY=83.0 ～ 184.0). MeixB was lower than the control group in group IV (U=72.0), and the differences were statistically significant (all P<0.05). MeixD and meixF showed consistent expression, with higher expression in groups III，IV and V compared to the control group (UmeixD =34.0 ～ 102.0，UmeixF=65.0 ～ 113.0). MeixX was expressed higher in groups II, IV and V compared to the control group (U=164.0，58.0，111.0), while oprD was only expressed lower in group III than in the control group (U=140.0), with statistically significant differences (all P<0.05). Although the expression of oprD in groups II, IV and V was lower than that in the control group, the differences were not statistically significant (U=381.0，102.0，144.0，all P>0.05). Conclusion ExCD, mexEF and mexXY are the main membrane protein combinations of CRPA efflux pumps in Kunming area. Upregulation of mexD, E, F, X, and Y membrane protein expression enhanced efflux. The correlation between mexAB oprM efflux pump and carbapenem resistance in CRPA in this area was low. The low expression of oprD played a role in the efflux mechanism in strains that do not produce β-lactase, but there was no significant difference in low expression in enzyme producing strains.

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备注/Memo

备注/Memo:
基金项目：昆明市卫生健康委员会卫生科研项目（2022-11-01-022）。
作者简介：卢赞（1978-），男，学士，副主任技师，研究方向：临床微生物检验，E-mail：luzan_km@163.com。
更新日期/Last Update: 2025-01-15