Study of <i>OprD2</i> gene polymorphism in imipenem-resistant <i>Pseucionionas aeruginosa</i>_West China Medical Journal

Authors：

BAI Tingting , WANG Dan , FAN Yingzi , YANG Xianggui , ZHOU Qin ,  XU Ying

Department of Clinical Laboratory, the First Affiliated Hospital of Chengdu Medical College, Chengdu, Sichuan 610500, P. R. China;

Corresponding author：

XU Ying, Email: yingxu@cmc.edu.cn

Keywords：

Pseudomonas aeruginosa; Imipenem; OprD2 gene; Drug-resistant mechanism

DOI：

10.7507/1002-0179.202006367

Video：

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Objective To explore the relationship between imipenem-resistant Pseudomonas aeruginosa (IRPA) and outer membrane porin protein OprD2 gene mutation.Methods IRPA strains (n=30) and imipenem-sensitive Pseudomonas aeruginosa strains (n=30) isolated from the clinical specimens in the First Affiliated Hospital of Chengdu Medical College from December 2018 to December 2019 were collected. Bacteria identification and drug sensitivity experiments were performed by VITEK-2 Compact combined with Kirby-Bauer method. Quantitative real-time polymerase chain reaction was used to detect the expression levels of OprD2 gene in the imipenem-resistant group and the imipenem-sensitive group, and then the strains with decreased expression were sequenced.Results The expression level of OprD2 gene in the imipenem-resistant group was significantly lower than that in the imipenem-sensitive group (P=0.048). Compared with the X63152 sequence, all the 11 Pseudomonas aeruginosa strains with significantly decreased OprD2 expression carried genetic variation, which occurred in coding regions. The variation sites presented diversity. The missense mutation of c.308C→G, c.344A→C, c.379G→C, c.471G→C, c.508T→C, c.553G→C, c.556-558CCG→GGC and c.565-566TG→AC caused amino acid change in the loop L2 and L3 of OprD2 porin, which affected the binding to imipenem. In addition, the mutations at 127, 169-171, 175, 177, 604, 628-630, 688, 719, 785, 826, 828, 842-843, 886, 901, 928-930, 934, 936, 944-945, 1039, 1041 and 1274 all resulted in the changes of amino acid. We also detected a deletion (c.1114-1115delAT) and other nonsense mutations. Large fragment deletion of OprD2 gene occurred in Strain 12. Conclusions The mutation and deletion of OprD2 gene can reduce the expression lever of OprD2 gene, leading to the resistance to imipenem of Pseudomonas aeruginosa. The variation of OprD2 gene of IRPA from clinical strains is diverse.

Citation： BAI Tingting, WANG Dan, FAN Yingzi, YANG Xianggui, ZHOU Qin, XU Ying. Study of OprD2 gene polymorphism in imipenem-resistant Pseucionionas aeruginosa. West China Medical Journal, 2020, 35(8): 924-929. doi: 10.7507/1002-0179.202006367 Copy

1.	Karampatakis T, Antachopoulos C, Tsakris A, <italic>et al</italic>. Molecular epidemiology of carbapenem-resistant <italic>Pseudomonas aeruginosa</italic> in an endemic area: comparison with global data. Eur J Clin Microbiol Infect Dis, 2018, 37(7): 1211-1220.
2.	陈约慧, 应芙蓉, 詹玲玲, 等. 1543 株铜绿假单胞菌感染的临床分布与耐药性分析. 中国临床药理学杂志, 2018, 34(24): 2814-2817.
3.	国家卫生计生委合理用药专家委员会, 全国细菌耐药监测网. 2018 年全国细菌耐药监测报告. 中国合理用药探索, 2020, 17(1): 1-10.
4.	Yoneyama H, Nakae T. Mechanism of efficient elimination of protein-D2 in outer-membrane of imipenem-resistant <italic>Pseudomonas-aeruginosa</italic>. Antimicrob Agents Chemother, 1993, 37(11): 2385-2390.
5.	Fang ZL, Zhang LY, Huang YM, <italic>et al</italic>. <italic>OprD</italic> mutations and inactivation in imipenem-resistant <italic>Pseudomonas aeruginosa</italic> isolates from China. Infect Genet Evol, 2014, 21: 124-128.
6.	Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. CLSI supplement M100. 30th ed. Wayne, PA: Clinical and Laboratory Standards Institute, 2020.
7.	梁文, 李博, 董萌萌, 等. 铜绿假单胞菌对碳青霉烯类抗生素耐药机制研究进展. 中国抗生素杂志, 2013, 38(9): 641-646.
8.	Muderris T, Durmaz R, Ozdem B, <italic>et al</italic>. Role of efflux pump and OprD porin expression in carbapenem resistance of <italic>Pseudomonas aeruginosa</italic> clinical isolates. J Infect Dev Ctries, 2018, 12(1): 1-8.
9.	Li H, Luo YF, Williams BJ, <italic>et al</italic>. Structure and function of OprD protein in <italic>Pseudomonas aeruginosa</italic>: from antibiotic resistance to novel therapies. Int J Med Microbiol, 2012, 302(2): 63-68.
10.	Quinn JP, Dudek EJ, Divincenzo CA, <italic>et al</italic>. Emergence of resistance to imipenem during therapy for <italic>Pseudomonas aeruginosa</italic> infections. J Infect Dis, 1986, 154(2): 289-294.
11.	沈继录, 朱德妹, 吴卫红, 等. 碳青霉烯类抗生素耐药铜绿假单胞菌外膜孔蛋白 OprD2 的研究. 中国感染与化疗杂志, 2011, 11(4): 281-286.
12.	颜英俊, 喻华, 周忠华, 等. 亚胺培南耐药的铜绿假单胞菌中 <italic>oprD</italic> 基因突变研究. 中华检验医学杂志, 2009, 32(4): 451-454.
13.	Kim CH, Kang HY, Kim BR, <italic>et al</italic>. Mutational inactivation of OprD in carbapenem-resistant <italic>Pseudomonas aeruginosa</italic> isolates from Korean hospitals. J Microbiol, 2016, 54(1): 44-49.
14.	宋诗铎, 郑东钧, 张宇雯, 等. 临床分离耐亚胺培南铜绿假单胞菌染色体 <italic>oprD</italic> 基因突变的检测. 中华传染病杂志, 2003, 21(2): 144-145.
15.	李飞, 李丽娟, 梁德志, 等. 耐亚胺培南铜绿假单胞菌耐药特征及其耐药机制的研究. 分子诊断与治疗杂志, 2019, 11(2): 101-106, 140.
16.	Epp SF, Köhler T, Plésiat P, <italic>et al</italic>. C-terminal region of <italic>Pseudomonas aeruginosa</italic> outer membrane porin OprD modulates susceptibility to meropenem. Antimicrob Agents Chemother, 2001, 45(6): 1780-1787.
17.	席红利, 李娟, 杨丽娜, 等. 耐亚胺培南铜绿假单胞菌的耐药性特征及 <italic>oprD</italic> 基因突变分析. 基因组学与应用生物学, 2019, 38(12): 5684-5688.
18.	李方去, 刘洋, 杨锦红, 等. 儿科铜绿假单胞菌的 <italic>OprD</italic> 基因突变及表达对碳青霉烯类抗生素耐药的作用. 中国卫生检验杂志, 2011, 21(5): 1063-1066.

1. Karampatakis T, Antachopoulos C, Tsakris A, <italic>et al</italic>. Molecular epidemiology of carbapenem-resistant <italic>Pseudomonas aeruginosa</italic> in an endemic area: comparison with global data. Eur J Clin Microbiol Infect Dis, 2018, 37(7): 1211-1220.
2. 陈约慧, 应芙蓉, 詹玲玲, 等. 1543 株铜绿假单胞菌感染的临床分布与耐药性分析. 中国临床药理学杂志, 2018, 34(24): 2814-2817.
3. 国家卫生计生委合理用药专家委员会, 全国细菌耐药监测网. 2018 年全国细菌耐药监测报告. 中国合理用药探索, 2020, 17(1): 1-10.
4. Yoneyama H, Nakae T. Mechanism of efficient elimination of protein-D2 in outer-membrane of imipenem-resistant <italic>Pseudomonas-aeruginosa</italic>. Antimicrob Agents Chemother, 1993, 37(11): 2385-2390.
5. Fang ZL, Zhang LY, Huang YM, <italic>et al</italic>. <italic>OprD</italic> mutations and inactivation in imipenem-resistant <italic>Pseudomonas aeruginosa</italic> isolates from China. Infect Genet Evol, 2014, 21: 124-128.
6. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing. CLSI supplement M100. 30th ed. Wayne, PA: Clinical and Laboratory Standards Institute, 2020.
7. 梁文, 李博, 董萌萌, 等. 铜绿假单胞菌对碳青霉烯类抗生素耐药机制研究进展. 中国抗生素杂志, 2013, 38(9): 641-646.
8. Muderris T, Durmaz R, Ozdem B, <italic>et al</italic>. Role of efflux pump and OprD porin expression in carbapenem resistance of <italic>Pseudomonas aeruginosa</italic> clinical isolates. J Infect Dev Ctries, 2018, 12(1): 1-8.
9. Li H, Luo YF, Williams BJ, <italic>et al</italic>. Structure and function of OprD protein in <italic>Pseudomonas aeruginosa</italic>: from antibiotic resistance to novel therapies. Int J Med Microbiol, 2012, 302(2): 63-68.
10. Quinn JP, Dudek EJ, Divincenzo CA, <italic>et al</italic>. Emergence of resistance to imipenem during therapy for <italic>Pseudomonas aeruginosa</italic> infections. J Infect Dis, 1986, 154(2): 289-294.
11. 沈继录, 朱德妹, 吴卫红, 等. 碳青霉烯类抗生素耐药铜绿假单胞菌外膜孔蛋白 OprD2 的研究. 中国感染与化疗杂志, 2011, 11(4): 281-286.
12. 颜英俊, 喻华, 周忠华, 等. 亚胺培南耐药的铜绿假单胞菌中 <italic>oprD</italic> 基因突变研究. 中华检验医学杂志, 2009, 32(4): 451-454.
13. Kim CH, Kang HY, Kim BR, <italic>et al</italic>. Mutational inactivation of OprD in carbapenem-resistant <italic>Pseudomonas aeruginosa</italic> isolates from Korean hospitals. J Microbiol, 2016, 54(1): 44-49.
14. 宋诗铎, 郑东钧, 张宇雯, 等. 临床分离耐亚胺培南铜绿假单胞菌染色体 <italic>oprD</italic> 基因突变的检测. 中华传染病杂志, 2003, 21(2): 144-145.
15. 李飞, 李丽娟, 梁德志, 等. 耐亚胺培南铜绿假单胞菌耐药特征及其耐药机制的研究. 分子诊断与治疗杂志, 2019, 11(2): 101-106, 140.
16. Epp SF, Köhler T, Plésiat P, <italic>et al</italic>. C-terminal region of <italic>Pseudomonas aeruginosa</italic> outer membrane porin OprD modulates susceptibility to meropenem. Antimicrob Agents Chemother, 2001, 45(6): 1780-1787.
17. 席红利, 李娟, 杨丽娜, 等. 耐亚胺培南铜绿假单胞菌的耐药性特征及 <italic>oprD</italic> 基因突变分析. 基因组学与应用生物学, 2019, 38(12): 5684-5688.
18. 李方去, 刘洋, 杨锦红, 等. 儿科铜绿假单胞菌的 <italic>OprD</italic> 基因突变及表达对碳青霉烯类抗生素耐药的作用. 中国卫生检验杂志, 2011, 21(5): 1063-1066.

West China Medical Journal

Study of OprD2 gene polymorphism in imipenem-resistant Pseucionionas aeruginosa

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