亚胺培南耐药铜绿假单胞菌产碳青霉烯酶耐药机制及其分子流行病学特征研究
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摘要
目的:检测和分析亚胺培南耐药铜绿假单胞菌(IRPA)碳青霉烯酶耐药机制,研究其分子流行病学特征,为防治院内IRPA感染提供参考。方法:抽取2016—2017年间临床分离出的IRPA菌株80株资料(产/非产碳青霉烯酶IRPA菌株,分别为32株和48株),分析采用改良碳青霉烯灭活试验(mCIM法)和改良Hodge法和聚合酶链反应(PCR)法检测碳青霉烯酶基因,其显示阳性者进行测序比较产/非产碳青霉烯酶IRPA菌株的耐药谱差异性,以及分析对携带不同种类碳青霉烯酶菌株的耐药及流行病学特征。结果:分离出的80株IRPA中,其中32株(40.00%)mCIM表型阳性,15株(18.75%)为改良Hodge试验阳性,PCR检测结果显示VIM-2基因阳性13株(16.25%)、KPC-2基因阳性19株(23.75%),NDM及OXA-48基因均为阴性;产碳青霉烯酶IRPA菌株对头孢他啶、头孢吡肟、哌拉西林-他唑巴坦、头孢哌酮-舒巴坦和阿米卡星的耐药率分别为87.50%、87.50%、68.75%、68.75%和68.75%显著高于非产碳青霉烯酶IRPA菌株分别为37.50%、35.42%、22.92%、22.92%和35.42%(P<0.05);MLST及PFGE分型结果显示IRPA菌株11个基因型,以ST235/A、ST244/D及ST639/K型为主,其中产酶IRPA菌株主要流行在ST235/A型、ST235/G型、ST244/D型、ST639/K型及ST1029/G型中。结论:本地区分离出的IRPA菌株以产VIM-2、KPC-2型MBL为主,其对临床常用的多种抗菌药物具有高度耐药性,应加强对产酶菌株的检测和监控,防止该菌株对抗菌药物的耐药性传播和流行。
Objective: To detect and analyze the mechanism of carbapenem resistance by imipenem resistant P. Aeruginosa(IRPA), and to study its molecular epidemiological characteristics, so as to provide a reference for the prevention and treatment of hospital IRPA infection. Methods: Clinical data from 2016 to 2017 IRPA strains in 80 strains were extracted and strains isolated(producting/non producing IRPA strains, 32 strains and 48 strains, respectively); analyzing of modified carbon black alkene inactivated test method(mCIM) and the method of modified Hodge and polymerase chain reaction(PCR) method to detect the carbapenem enzyme gene; its sequencing showed that positive was to produce/non carbon penicillium enzyme drug-resistant spectrum difference of IRPA strains, as well as analyzing different types of carbon penicillium enzyme resistant and epidemiological characteristics of the strain. Results: Of the 80 IRPA strains isolated, 32(40.00%) were mCIM phenotype positive, and 15(18.75%) were modified Hodge positive; the PCR results showed that 13(16.25%) were VIM-2 gene positive, 19(23.75%) were kpc-2 gene positive, and both NDM and oxa-48 genes were negative. The drug resistance rates of IRPA strains producing carbapenem to ceftazidime, cefepime, piperacilin-tazobactam, cefoperazone, sulbactam and amikacin were 87.50%, 87.50%, 68.75%, 68.75%, and 68.75%, respectively, which were significantly higher than those of non-carbapene-producing IRPA strains(37.50%, 35.42%, 22.92%, 22.92%, and35.42%, respectively)(P<0.05). The results of MLST and PFGE typing showed that IRPA strains had 11 genotypes, mainly ST235/A, ST244/D and ST639/K, among which the enzyme-producing IRPA strains were mainly prevalent in ST235/A, ST235/G, ST244/D, ST639/K and ST1029/G. Conclusions: IRPA strains isolated in this region mainly produce VIM-2 and KPC-2 MBL, which were highly resistant to a variety of commonly used clinical antimicrobial agents. The detection and monitoring of enzyme-producing strains should be strengthened to prevent the spreading and prevalence of antimicrobial resistance of these strains.
Objective: To detect and analyze the mechanism of carbapenem resistance by imipenem resistant P. Aeruginosa(IRPA), and to study its molecular epidemiological characteristics, so as to provide a reference for the prevention and treatment of hospital IRPA infection. Methods: Clinical data from 2016 to 2017 IRPA strains in 80 strains were extracted and strains isolated(producting/non producing IRPA strains, 32 strains and 48 strains, respectively); analyzing of modified carbon black alkene inactivated test method(mCIM) and the method of modified Hodge and polymerase chain reaction(PCR) method to detect the carbapenem enzyme gene; its sequencing showed that positive was to produce/non carbon penicillium enzyme drug-resistant spectrum difference of IRPA strains, as well as analyzing different types of carbon penicillium enzyme resistant and epidemiological characteristics of the strain. Results: Of the 80 IRPA strains isolated, 32(40.00%) were mCIM phenotype positive, and 15(18.75%) were modified Hodge positive; the PCR results showed that 13(16.25%) were VIM-2 gene positive, 19(23.75%) were kpc-2 gene positive, and both NDM and oxa-48 genes were negative. The drug resistance rates of IRPA strains producing carbapenem to ceftazidime, cefepime, piperacilin-tazobactam, cefoperazone, sulbactam and amikacin were 87.50%, 87.50%, 68.75%, 68.75%, and 68.75%, respectively, which were significantly higher than those of non-carbapene-producing IRPA strains(37.50%, 35.42%, 22.92%, 22.92%, and35.42%, respectively)(P<0.05). The results of MLST and PFGE typing showed that IRPA strains had 11 genotypes, mainly ST235/A, ST244/D and ST639/K, among which the enzyme-producing IRPA strains were mainly prevalent in ST235/A, ST235/G, ST244/D, ST639/K and ST1029/G. Conclusions: IRPA strains isolated in this region mainly produce VIM-2 and KPC-2 MBL, which were highly resistant to a variety of commonly used clinical antimicrobial agents. The detection and monitoring of enzyme-producing strains should be strengthened to prevent the spreading and prevalence of antimicrobial resistance of these strains.
引文
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[15]姜波,吴金英,伊茂礼,等.烟台地区产金属-内β-酰胺酶铜绿假单胞菌的耐药性分析及基因检测[J].国际检验医学杂志,2009,30(7),641-644.
[16]徐令清,王欢,温伟洪,等.铜绿假单胞菌多位点序列分析[J].中华医院感染学杂志,2017,27(1),35-37.
[17]Rajbharan YD,AJürgen B,BRoger L,et al.Optimization of synergistic combination regimens against carbapenemand aminoglycoside-resistant clinical Pseudomonas aeruginosa Isolates via mechanism-based pharmacokinetic/pharmacodynamic modeling[J].Antimicrob Agents and Chemoth,2017,61(1):e01011-16.
[18]Jisun K,Bora S,Chulwoo P,et al.Indole-induced activities ofβ-lactamase and efflux pump confer ampicillin resistance in Pseudomonas putida KT2440[J].Frontiers in Microbiol,2017,(8):433-447.
[2]Luciana CC,Stephanie GC,Natacha M,et al.Mechanisms of carbapenem resistance in endemic Pseudomonas aeruginosa isolates after an SPM-1 metallo-β-lactamase producing strain subsided in an intensive care unit of a teaching hospital in Brazil[J].Mem Inst Oswaldo Cruz,Rio de Janeiro,2016,111(9):551-558.
[3]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.
[4]Cai SQ,Chen YQ,Song DZ,et al.Study on the resistance mechanism via outer membrane protein OprD2 and metalβ-lactamase expression in the cell wall of Pseudomonas aeruginosa[J].Experiment and Therap Med,2016,12:2869-2872.
[5]郝士卿,杨浩宁,尤文文,等.372株铜绿假单胞菌的临床分布特点及耐药性分析[J].临床合理用药,2017,10(1):154-157.
[6]曲彩红,江先合.175株铜绿假单胞菌的检测情况及耐药性分析[J].2008第十一次全国临床药理学学术大会,72-73.
[7]牛文娟.铜绿假单胞菌对14种抗菌药物的多重耐药分析[J].中国医药导报,2010,7(1),181.
[8]黄学忠,林佩佩,陈晓飞.痰标本铜绿假单胞菌224株5年耐药变迁[J].检验医学与临床,2013,10(3),560-563.
[9]应春妹,应骏,罗柳林,等.耐亚胺培南铜绿假单胞菌耐药机制研究[J].中国感染与化疗杂志,2008,20(7),300-303.
[10]朱星成,夏鹏.铜绿假单胞菌的临床分布及耐药分析[J].中华临床医师杂志,2016,(10):54-55.
[11]潘婉仪,黄宪章,杨洁,等.广州地区产β-内酰胺酶铜绿假单胞菌的流行病学研究[J].南方医科大学学报,2010,30(8):1893-1895.
[13]刘建雷,仇杭佳.铜绿假单胞菌医院感染特点及耐药分析[J].国际检验医学杂志,2010,31(12),1436-1437.
[13]姚蓓,张丽丽.重庆某二甲医院铜绿假单胞菌耐药分析[J].国际检验医学杂志,2016,37(11),3046-3047.
[14]杜飞.铜绿假单胞菌耐药分析与基因研究[J].中国药物与临床,2013,13(3),400-401.
[15]姜波,吴金英,伊茂礼,等.烟台地区产金属-内β-酰胺酶铜绿假单胞菌的耐药性分析及基因检测[J].国际检验医学杂志,2009,30(7),641-644.
[16]徐令清,王欢,温伟洪,等.铜绿假单胞菌多位点序列分析[J].中华医院感染学杂志,2017,27(1),35-37.
[17]Rajbharan YD,AJürgen B,BRoger L,et al.Optimization of synergistic combination regimens against carbapenemand aminoglycoside-resistant clinical Pseudomonas aeruginosa Isolates via mechanism-based pharmacokinetic/pharmacodynamic modeling[J].Antimicrob Agents and Chemoth,2017,61(1):e01011-16.
[18]Jisun K,Bora S,Chulwoo P,et al.Indole-induced activities ofβ-lactamase and efflux pump confer ampicillin resistance in Pseudomonas putida KT2440[J].Frontiers in Microbiol,2017,(8):433-447.