耐亚胺培南铜绿假单胞菌耐药特征及产金属β-内酰胺酶基因型分析
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摘要
目的研究耐亚胺培南铜绿假单胞菌对常用抗生素耐药性以及其产金属β-内酰胺酶(MBL)的基因型。方法收集2016-2017年东莞市人民医院以及东莞市东华医院2家医院的110株耐亚胺培南铜绿假单胞菌临床分离株,采用VITEK2系统分析临床菌株耐药性;采用组合纸片法对亚胺培南耐药菌株进行MBL表型检测,同时采用PCR法检测MBL相关基因(包括IMP型、VIM型、SPM型、SIM型、GIM型和NDM型);比较分析产与不产MBL的耐亚胺培南铜绿假单胞菌耐药谱差异。结果耐亚胺培南铜绿假单胞菌对阿米卡星的耐药率为10.00%,对妥布霉素、庆大霉素、头孢吡肟、哌拉西林/他唑巴坦的耐药率在17.27%～21.82%之间,对头孢他啶、环丙沙星和左氧氟沙星的耐药率为26.36%～31.82%,对其余β-内酰胺类、氟喹诺酮类、四环素、磺胺类抗生素耐药严重(74.54%～100.00%)。110株耐亚胺培南铜绿单胞菌中筛选出18株(16.36%)MBL表型阳性的菌株,用PCR法也检测出18株MBL基因阳性的菌株(16.36%),其中9株检出是IMP-25(8.18%),8株检出是VIM-2(7.27%),4株检出是SIM-2(3.64%),有3株同时检出VIM-2和SIM-2,其他基因型均阴性。产MBL菌株对所有药物的耐药率高于非产MBL菌株。结论产金属β-内酰胺酶在铜绿假单胞菌对亚胺培南耐药中发挥重要作用,酶基因型主要为IMP-25、VIM-2和SIM-2。
Objective To study the drug resistance and metallo-β-lactamase gene(MBL) of clinical isolates of Pseudomonas aeruginosa. Methods 110 imipenem-resistance Pseudomonas aeruginosa clinical isolates were collected from 2 hospitals of Dongguan between 2016 and 2017, and antibiotic resistance was analyzed using VITEK2 system. With the combination of disk diffusion method the metallo-β-lactamases( MBL) phenotype of imipenem-resistant isolates was detected. PCR was performed to amplify gene MBL. The antibiotic resistance of MBL-producing isolates(IMP, VIM,SPM,SIM, GIM, NDM) of imipenem resistant Pseudomonas aeruginosa was compared with that of non MBL Producing isolates.Results Impenem-resistant Pseudomonas aeruginosa had the lowest resistance rate to amikacin( 10.00%), its resistance to tobramycin, gentamycin, cefepime, piperacillin/tazobactam was 17.27%～21.82%and its resistance to ceftazidime,ciprofloxacin and levofloxacin was 26.36%～31.82% and other antibacterial drugs of β-lactamase, fluoroquinolones,tetracycline and sulfonamides are seriously resistance(74.54%～100.00%). 18 strains of MBL positive strain(16.36%)were detected in 110 strains of imipenem-resistant Pseudomonas aeruginosa, and 9( 8.18%) tested positive for IMP-25, 8(7.27%) tested positive for VIM-2, 4(3.64%)tested positive for SIM-2, 3 tested for both VIM-2 and SIM-2. Other MBL genes were not detected. The resistance rates to all antibiotics of MBL-producing isolates were higher than those of non MBLproducing isolates. Conclusion Producing metallo-β-lactamase was one of the main mechanisms of drug resistance of Pseudomonas aeruginosa to imipenem, and the main genotypes were IMP-25, VIM-2 and SIM-2.
Objective To study the drug resistance and metallo-β-lactamase gene(MBL) of clinical isolates of Pseudomonas aeruginosa. Methods 110 imipenem-resistance Pseudomonas aeruginosa clinical isolates were collected from 2 hospitals of Dongguan between 2016 and 2017, and antibiotic resistance was analyzed using VITEK2 system. With the combination of disk diffusion method the metallo-β-lactamases( MBL) phenotype of imipenem-resistant isolates was detected. PCR was performed to amplify gene MBL. The antibiotic resistance of MBL-producing isolates(IMP, VIM,SPM,SIM, GIM, NDM) of imipenem resistant Pseudomonas aeruginosa was compared with that of non MBL Producing isolates.Results Impenem-resistant Pseudomonas aeruginosa had the lowest resistance rate to amikacin( 10.00%), its resistance to tobramycin, gentamycin, cefepime, piperacillin/tazobactam was 17.27%～21.82%and its resistance to ceftazidime,ciprofloxacin and levofloxacin was 26.36%～31.82% and other antibacterial drugs of β-lactamase, fluoroquinolones,tetracycline and sulfonamides are seriously resistance(74.54%～100.00%). 18 strains of MBL positive strain(16.36%)were detected in 110 strains of imipenem-resistant Pseudomonas aeruginosa, and 9( 8.18%) tested positive for IMP-25, 8(7.27%) tested positive for VIM-2, 4(3.64%)tested positive for SIM-2, 3 tested for both VIM-2 and SIM-2. Other MBL genes were not detected. The resistance rates to all antibiotics of MBL-producing isolates were higher than those of non MBLproducing isolates. Conclusion Producing metallo-β-lactamase was one of the main mechanisms of drug resistance of Pseudomonas aeruginosa to imipenem, and the main genotypes were IMP-25, VIM-2 and SIM-2.
引文
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[13]闫玉兰,郭世辉,李萌,等.耐亚胺培南铜绿假单胞菌的耐药机制研究[J].中华医院感染学杂志,2014,24(15):3650-3652.
[14]方小龙,王莉平,林立鹏,等.耐亚胺培南铜绿假单胞菌金属β-内酰胺酶基因及耐消毒剂基因研究[J].中国热带医学,2018,18(2):107-115.
[15]唐慧琴,陈定强,吴爱武,等.耐亚胺培南铜绿假单胞菌的金属β-内酰胺酶检测[J].中国卫生检验杂志,2014,24(6):831-833.
[16]余广超,徐霖,袁广卿,等.亚胺培南耐药铜绿假单胞菌的金属β-内酰胺酶检测[J].中国抗生素杂志,2011,36(3):223-227.
[17]Mohanam L,Menon T.Coexistence of metallo-beta-lactamaseencoding genes in Pseudomonas aeruginosa[J].Indian J Med Res,2018,29(16):46-52.
[18]Bellés A,Bueno J,Rojo-Bezares B,et al.Characterisation of VIM-2 producing Pseudomonas aeruginosa isolates from lower tract respiratory infections in a Spanish hospital[J].Eur J Clin Microbiol Infect Dis,2018,37(10):1847-1856.
[19]万强,薄志坚,陈杨,等.大连地区耐亚胺培南铜绿假单胞菌金属β-内酰胺酶检测及耐药性分析[J].中国微生态学杂志,2016,28(4):405-408.
[20]何力志,黄露萍,李志芳,等.耐亚胺培南铜绿假单胞菌金属β-内酰胺酶的检测及耐药性分析[J].检验医学,2013,28(5):353-356.
[2]Liu Q,Li X,Li W,et al.Influence of carbapenem resistance on mortality of patients with Pseudomonas aeruginosa infection:Ameta-analysis[J].Sci Rep,2015,5:11715.
[3]胡付品,郭燕,朱德妹,等.2016年中国CHINET细菌耐药性监测[J].中国感染与化疗杂志,2017,17(5):481-491.
[4]曾为伦,吴海鸥,章如玲,等.多重耐药铜绿假单胞菌的相关耐药机制研究[J].中国抗生素杂志,2016,41(5):377-381.
[5]陈宏标,苏丽贤,吴紫娟,等.亚胺培南耐药PA的基因型及耐药机制研究[J].中国热带医学,2015,5(8):928-931.
[6]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[J].分子诊断与治疗杂志,2019,11(2):101-106,140.
[7]佘丹阳.严峻耐药形势下碳青霉烯类抗生素的临床应用[J].中国感染与化疗杂志,2013,13(4):317-318.
[8]龚映,傅跃光.多重耐药铜绿假单胞菌耐药机制研究进展[J].儿科药学杂志,2015,21(8):58-61.
[9]Arakawa Y,Shibata N,Shibayama K,et al.Convenient test for screening metallo-beta-lactamase-producing gram-negative bacteria by using thiol compounds[J].J Clin Microbiol,2000,38(1):40-43.
[10]Ellington MJ,Kistler J,Livermore DM,et al.Multiplex PCR for rapid detection of genes encoding acquired metallo-β-lactamases[J].J Antimicrob Chemother,2007,59(2):321-322.
[11]Laurent P,Gunturu R,Sandrine B,et al.Detection of NDM-1-Producing Klebsiella pneumoniae in Kenya[J].Antimicrob Agents Chemother,2011,55(2):934-936.
[12]Swayne R,Ellington MJ,Curran MD,et al.Utility of a novel multiplex TaqMan PCR assay for metallo-β-lactamase genes plus other TaqMan assays in detecting genes encoding serine carbapenemases and clinically significant extended-spectrumβ-lactamases[J].Int J Antimicrob Agents,2013,42(4):352-356.
[13]闫玉兰,郭世辉,李萌,等.耐亚胺培南铜绿假单胞菌的耐药机制研究[J].中华医院感染学杂志,2014,24(15):3650-3652.
[14]方小龙,王莉平,林立鹏,等.耐亚胺培南铜绿假单胞菌金属β-内酰胺酶基因及耐消毒剂基因研究[J].中国热带医学,2018,18(2):107-115.
[15]唐慧琴,陈定强,吴爱武,等.耐亚胺培南铜绿假单胞菌的金属β-内酰胺酶检测[J].中国卫生检验杂志,2014,24(6):831-833.
[16]余广超,徐霖,袁广卿,等.亚胺培南耐药铜绿假单胞菌的金属β-内酰胺酶检测[J].中国抗生素杂志,2011,36(3):223-227.
[17]Mohanam L,Menon T.Coexistence of metallo-beta-lactamaseencoding genes in Pseudomonas aeruginosa[J].Indian J Med Res,2018,29(16):46-52.
[18]Bellés A,Bueno J,Rojo-Bezares B,et al.Characterisation of VIM-2 producing Pseudomonas aeruginosa isolates from lower tract respiratory infections in a Spanish hospital[J].Eur J Clin Microbiol Infect Dis,2018,37(10):1847-1856.
[19]万强,薄志坚,陈杨,等.大连地区耐亚胺培南铜绿假单胞菌金属β-内酰胺酶检测及耐药性分析[J].中国微生态学杂志,2016,28(4):405-408.
[20]何力志,黄露萍,李志芳,等.耐亚胺培南铜绿假单胞菌金属β-内酰胺酶的检测及耐药性分析[J].检验医学,2013,28(5):353-356.