耐碳青霉烯类肺炎克雷伯菌耐药性及碳青霉烯类失活法检测分析
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摘要
目的分析临床分离的耐碳青霉烯类肺炎克雷伯菌(CRKP)对临床常用药物的耐药情况及碳青霉烯类失活法(CIM)、改良Hodge试验(MHT)对产碳青霉烯酶的筛选能力。方法使用聚合酶链式反应(PCR)扩增和琼脂糖凝胶电泳的方法检测菌株携带碳青霉烯酶相关基因结果为标准,评估CIM和MHT对医院2016年1月-2017年9月临床分离的非重复30株CRKP及40株敏感菌株的碳青霉烯酶(CSKP)表型筛选能力。结果 30株CRKP对替加环素、阿米卡星、磺胺甲噁唑/甲氧苄啶的耐药率分别为0、13.33%、16.67%,对头孢哌酮/舒巴坦、哌拉西林/他唑巴坦和美罗培南耐药率分别为86.67%、93.33%、93.33%,对其他9种β-内酰胺类抗菌药物的耐药率为100.00%;除磺胺甲噁唑/甲氧苄啶、头孢唑林及替加环素外,CRKP对其他抗菌药物的耐药性较CSKP差异有统计学意义(P<0.05)。PCR结果表明30株CRKP均携带碳青霉烯酶基因;CIM和MHT灵敏度、特异度、阳性预测值和阴性预测值分别为100.00%、100.00%、100.00%和100.00%;96.67%、100.00%、100.00%和97.56%。结论 CRKP对常用抗菌药物表现出较高耐药性,碳青霉烯酶的产生是主要耐药机制,提示临床应加强抗菌药物的管理;与PCR相比,MHT和CIM均具有较高的灵敏度、特异度和预测值,可在常规实验室开展,CIM具有结果更准确、更易于观察、结果读取时间短等优势。
OBJECTIVE To evaluate the antimicrobial susceptibility of carbapenem-resistant Klebsiella pneumoniae(CRKP),as well as the ability of carbapenem inactivation method(CIM)and modified Hodge test(MHT)in detecting carbapenemase production.METHODS PCR and agarose gel electrophoresis were used to detect the genes of carbapenemase in K pneumoniae strains,and the results were utilized as the standard of the phenotype screening capacity of CIM and MHT test for 30 non-repetitive clinical carbapenem-resistant K pneumoniae strains and40 susceptible strains(CSKP)collected in the hospital from Jan.2016 to Sep.2017.RESULTS The resistance rates of 30 strains of CRKP to tigecycline,amikacin and Trimethoprim/sulfamethoxazole were 0,13.33%,16.07%,respectively.The resistance rate to cefoperazone/sulbactam was 86.67%,piperacillin/tazobactam was 93.33%,meropenem was 93.33%,and 9 other kinds ofβ-lactam antibiotics were 100.00%.Except for trimethoprim/sulfamethoxazole,cefazolin,and tigecycline,CRKP had significantly higher resistance to other antibacterial agents than CSKP(P<0.05).The PCR results showed that all 30 strains of CRKP carried carbapenemase genes.The sensitivity,specificity,positive predictive value and negative predictive value of CIM and MHT were 100.00%,100.00%,100.00%and 100.00%;96.67%,100.00%,100.00%and 97.56%,respectively.CONCLUSIONCRKP showed high resistance to most of the common antimicrobial agents,and the production of carbapenemases is the major resistance mechanisms,suggesting that the clinicians should strengthen the monitoring of antibiotics use.Compared with PCR,MHT and CIM had a higher sensitivity,specificity and predictive value,and they can be carried out in conventional laboratories.CIM had the advantages of more accurate results,easier observation,and short reading time.
OBJECTIVE To evaluate the antimicrobial susceptibility of carbapenem-resistant Klebsiella pneumoniae(CRKP),as well as the ability of carbapenem inactivation method(CIM)and modified Hodge test(MHT)in detecting carbapenemase production.METHODS PCR and agarose gel electrophoresis were used to detect the genes of carbapenemase in K pneumoniae strains,and the results were utilized as the standard of the phenotype screening capacity of CIM and MHT test for 30 non-repetitive clinical carbapenem-resistant K pneumoniae strains and40 susceptible strains(CSKP)collected in the hospital from Jan.2016 to Sep.2017.RESULTS The resistance rates of 30 strains of CRKP to tigecycline,amikacin and Trimethoprim/sulfamethoxazole were 0,13.33%,16.07%,respectively.The resistance rate to cefoperazone/sulbactam was 86.67%,piperacillin/tazobactam was 93.33%,meropenem was 93.33%,and 9 other kinds ofβ-lactam antibiotics were 100.00%.Except for trimethoprim/sulfamethoxazole,cefazolin,and tigecycline,CRKP had significantly higher resistance to other antibacterial agents than CSKP(P<0.05).The PCR results showed that all 30 strains of CRKP carried carbapenemase genes.The sensitivity,specificity,positive predictive value and negative predictive value of CIM and MHT were 100.00%,100.00%,100.00%and 100.00%;96.67%,100.00%,100.00%and 97.56%,respectively.CONCLUSIONCRKP showed high resistance to most of the common antimicrobial agents,and the production of carbapenemases is the major resistance mechanisms,suggesting that the clinicians should strengthen the monitoring of antibiotics use.Compared with PCR,MHT and CIM had a higher sensitivity,specificity and predictive value,and they can be carried out in conventional laboratories.CIM had the advantages of more accurate results,easier observation,and short reading time.
引文
[1] Logan LK,Weinstein RA.The epidemiology of carbapenem-resistant Enterobacteriaceae:the impact and evolution of a global menace[J].J Infect Dis,2017,215(suppl_1):S28-S36.
[2]胡付品.2005-2014年CHINET中国细菌耐药性监测网5种重要临床分离菌的耐药性变迁[J].中国感染与化疗杂志,2017,17(1):93-99.
[3] Goodman KE,Simner PJ,Tamma PD,et al.Infection control implications of heterogeneous resistance mechanisms in carbapenem-resistant Enterobacteriaceae(CRE)[J].Expert Rev Anti Infect Ther,2016,14(1):95-108.
[4] Wayne,PA.Performance standards for antimicrobial susceptibility testing:twenty-th information supplement[J].CLSI document M100-S20,2010,30(1):1-53.
[5] Sun Y,Li M,Chen L,et al.Prevalence and molecular characterization of carbapenemase-producing gram-negative bacteria from a university hospital in China[J].Infect Dis,2016,48(2):138-146.
[6]周静芳,凌勇,刘伟江,等.耐碳青霉烯类肠杆菌科细菌的感染特征和危险因素分析[J].中华医院感染学杂志,2017,27(1):16-19.
[7] Adler A,Bendalak M,Chmelnitsky I,et al.Effect of resistance mechanisms on the inoculum effect of carbapenem in Klebsiella pneumoniae isolates with borderline carbapenem resistance[J].Antimicrob Agents Chemother,2015,59(8):5014-5017.
[8] Zhong X,Xu H,Chen D,et al.First emergence of acrAB and oqxAB mediated tigecycline resistance in clinical isolates of Klebsiella pneumoniae pre-dating the use of tigecycline in a Chinese hospital[J].PLoS One,2014,9(12):e115185.
[9] Bi W,Liu H,Dunstan RA,et al.Extensively drug-resistant Klebsiella pneumoniae causing nosocomial bloodstream infections in China:molecular investigation of antibiotic resistance determinants,informing therapy,and clinical outcomes[J].Front Microbiol,2017,8:1230.
[10] Yamada K,Kashiwa M,Arai K,et al.Comparison of the modified-hodge test,Carba NP test,and carbapenem inactivation method as screening methods for carbapenemase-producing Enterobacteriaceae[J].J Microbiol Methods,2016,128(9):48-51.
[11] van der Zwaluw K,de Haan A,Pluister GN,et al.The carbapenem inactivation method(CIM),a simple and low-cost alternative for the Carba NP test to assess phenotypic carbapenemase activity in gram-negative rods[J].PLoS One,2015,10(3):e0123690.
[12] Tijet N,Patel SN,Melano RG.Detection of carbapenemase activity in Enterobacteriaceae:comparison of the carbapenem inactivation method versus the Carba NP test[J].J Antimicrob Chemother,2016,71(1):274-276.
[13] Aguirre-Quinonero A,Cano ME,Gamal D,et al.Evaluation of the carbapenem inactivation method(CIM)for detecting carbapenemase activity in Enterobacteria[J].Diagn Microbiol Infect Dis,2017,88(3):214-218.
[14] Tamma PD,Huang Y,Opene BN,et al.Determining the optimal carbapenem MIC that distinguishes carbapenemaseproducing and non-carbapenemase-producing carbapenem-resistant Enterobacteriaceae[J].Antimicrob Agents Chemother,2016,60(10):6425-6429.
[2]胡付品.2005-2014年CHINET中国细菌耐药性监测网5种重要临床分离菌的耐药性变迁[J].中国感染与化疗杂志,2017,17(1):93-99.
[3] Goodman KE,Simner PJ,Tamma PD,et al.Infection control implications of heterogeneous resistance mechanisms in carbapenem-resistant Enterobacteriaceae(CRE)[J].Expert Rev Anti Infect Ther,2016,14(1):95-108.
[4] Wayne,PA.Performance standards for antimicrobial susceptibility testing:twenty-th information supplement[J].CLSI document M100-S20,2010,30(1):1-53.
[5] Sun Y,Li M,Chen L,et al.Prevalence and molecular characterization of carbapenemase-producing gram-negative bacteria from a university hospital in China[J].Infect Dis,2016,48(2):138-146.
[6]周静芳,凌勇,刘伟江,等.耐碳青霉烯类肠杆菌科细菌的感染特征和危险因素分析[J].中华医院感染学杂志,2017,27(1):16-19.
[7] Adler A,Bendalak M,Chmelnitsky I,et al.Effect of resistance mechanisms on the inoculum effect of carbapenem in Klebsiella pneumoniae isolates with borderline carbapenem resistance[J].Antimicrob Agents Chemother,2015,59(8):5014-5017.
[8] Zhong X,Xu H,Chen D,et al.First emergence of acrAB and oqxAB mediated tigecycline resistance in clinical isolates of Klebsiella pneumoniae pre-dating the use of tigecycline in a Chinese hospital[J].PLoS One,2014,9(12):e115185.
[9] Bi W,Liu H,Dunstan RA,et al.Extensively drug-resistant Klebsiella pneumoniae causing nosocomial bloodstream infections in China:molecular investigation of antibiotic resistance determinants,informing therapy,and clinical outcomes[J].Front Microbiol,2017,8:1230.
[10] Yamada K,Kashiwa M,Arai K,et al.Comparison of the modified-hodge test,Carba NP test,and carbapenem inactivation method as screening methods for carbapenemase-producing Enterobacteriaceae[J].J Microbiol Methods,2016,128(9):48-51.
[11] van der Zwaluw K,de Haan A,Pluister GN,et al.The carbapenem inactivation method(CIM),a simple and low-cost alternative for the Carba NP test to assess phenotypic carbapenemase activity in gram-negative rods[J].PLoS One,2015,10(3):e0123690.
[12] Tijet N,Patel SN,Melano RG.Detection of carbapenemase activity in Enterobacteriaceae:comparison of the carbapenem inactivation method versus the Carba NP test[J].J Antimicrob Chemother,2016,71(1):274-276.
[13] Aguirre-Quinonero A,Cano ME,Gamal D,et al.Evaluation of the carbapenem inactivation method(CIM)for detecting carbapenemase activity in Enterobacteria[J].Diagn Microbiol Infect Dis,2017,88(3):214-218.
[14] Tamma PD,Huang Y,Opene BN,et al.Determining the optimal carbapenem MIC that distinguishes carbapenemaseproducing and non-carbapenemase-producing carbapenem-resistant Enterobacteriaceae[J].Antimicrob Agents Chemother,2016,60(10):6425-6429.