mCIM与eCIM筛选肠杆菌科细菌碳青霉烯酶的效果评价
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摘要
目的评价改良碳青霉烯类失活法(modified carbapenem inactivation method,m CIM)和EDTA碳青霉烯类失活法(EDTAcarbapenem inactivation method,e CIM)对肠杆菌科细菌碳青霉烯酶表型的筛选能力。方法分别收集碳青霉烯类耐药和敏感肠杆菌科细菌102株和53株,采用m CIM和e CIM进行碳青霉烯酶表型筛选试验,PCR法检测blaKPC-2、blaNDM-1、blaIMP-4、blaVIM-1和blaOXA-48耐药基因,并对表型筛选试验结果与基因检测结果的一致性进行统计分析。结果 102株碳青霉烯类耐药肠杆菌科细菌中97株检出耐药基因,包括51株blaKPC-2基因、38株blaNDM-1基因、5株blaIMP-4基因以及3株同时携带blaKPC-2和blaNDM-1基因;m CIM检出阳性98株,阴性4株。98株m CIM阳性菌中,e CIM阳性46株,阴性52株。53株碳青霉烯类敏感肠杆菌科细菌耐药基因检测及m CIM试验均为阴性。m CIM试验筛选碳青霉烯类耐药肠杆菌科细菌碳青霉烯酶产生的敏感性为99.0%,特异性为96.6%,与PCR结果一致性Kappa值为0.959;e CIM筛选金属酶敏感性为93.5%,特异性为94.6%,Kappa值为0.881;e CIM筛选丝氨酸碳青霉烯酶敏感性为92.6%,特异性为95.8%,Kappa值为0.882。结论 m CIM试验与e CIM试验联合检测,不仅可以有效筛选碳青霉烯酶产酶株,而且可同时区分碳青霉烯酶类型,对流行病学调查研究及疾病治疗有重要意义。
Objective To evaluate the screening capacity of modified carbapenem inactivation method( m CIM) and EDTAcarbapenem inactivation method( e CIM) for phenotypic activity of carbapenemase in Enterobacteriaceae bacteria. Methods A total of102 isolates of carbapenem-resistant Enterobacteriaceae and 53 isolates of carbapenem-sensitive Enterobacteriaceae were selected and the carbapenemase activities were determined by m CIM and e CIM,respectively. Meanwhile,the carbapenem-resistant genes in Enterobacteriaceae,such as blaKPC-2,blaNDM-1,blaIMP-4,blaVIM-1 and blaOXA-48,were detected by PCR.The consistency of results between phenotypic screening tests and gene examinations was analyzed statistically. Results Of the 102 carbapenem-resistant Enterobacteriaceae strains,97 strains carrying positive drug-resistance gene were detected by PCR,including blaKPC-2 in 51 strains,blaNDM-1 in 38 strains,blaIMP-4 in 5 strains and both blaKPC-2 and blaNDM-1 in 3 strains. Of the 102 carbapenem-resistant Enterobacteriaceae strains,98 strains were detected to be positive phenotype by m CIM,while the other 4 strains were negative phenotype. Of the 98 m CIM positive strains,46 strains were e CIM positive and 52 strains were e CIM negative. Of the 53 carbapenem-sensitive strains,the results of PCR and m CIM were both negative. For screening of carbapenemase by m CIM,the sensitivity was 99.0% and the specificity was 96.6%,and the Kappa value for consistency with PCR results was 0.959. For screening metallo-beta-lactamase gene,the sensitivity of e CIM was 93.5%,the specificity was 94.6% and the Kappa value was 0.881. For screening class A carbapenemase gene,the sensitivity of e CIM was 92.6%,the specificity was 95.8%,and the Kappa value was 0.882. Conclusion The combined detections of m CIM and e CIM may effectively screen carbapenemase-producing strains in Enterobacteriaceae,and also distinguish the type of carbapenemase,which should be of great significance for epidemiological investigation and therapy of infectious diseases.
Objective To evaluate the screening capacity of modified carbapenem inactivation method( m CIM) and EDTAcarbapenem inactivation method( e CIM) for phenotypic activity of carbapenemase in Enterobacteriaceae bacteria. Methods A total of102 isolates of carbapenem-resistant Enterobacteriaceae and 53 isolates of carbapenem-sensitive Enterobacteriaceae were selected and the carbapenemase activities were determined by m CIM and e CIM,respectively. Meanwhile,the carbapenem-resistant genes in Enterobacteriaceae,such as blaKPC-2,blaNDM-1,blaIMP-4,blaVIM-1 and blaOXA-48,were detected by PCR.The consistency of results between phenotypic screening tests and gene examinations was analyzed statistically. Results Of the 102 carbapenem-resistant Enterobacteriaceae strains,97 strains carrying positive drug-resistance gene were detected by PCR,including blaKPC-2 in 51 strains,blaNDM-1 in 38 strains,blaIMP-4 in 5 strains and both blaKPC-2 and blaNDM-1 in 3 strains. Of the 102 carbapenem-resistant Enterobacteriaceae strains,98 strains were detected to be positive phenotype by m CIM,while the other 4 strains were negative phenotype. Of the 98 m CIM positive strains,46 strains were e CIM positive and 52 strains were e CIM negative. Of the 53 carbapenem-sensitive strains,the results of PCR and m CIM were both negative. For screening of carbapenemase by m CIM,the sensitivity was 99.0% and the specificity was 96.6%,and the Kappa value for consistency with PCR results was 0.959. For screening metallo-beta-lactamase gene,the sensitivity of e CIM was 93.5%,the specificity was 94.6% and the Kappa value was 0.881. For screening class A carbapenemase gene,the sensitivity of e CIM was 92.6%,the specificity was 95.8%,and the Kappa value was 0.882. Conclusion The combined detections of m CIM and e CIM may effectively screen carbapenemase-producing strains in Enterobacteriaceae,and also distinguish the type of carbapenemase,which should be of great significance for epidemiological investigation and therapy of infectious diseases.
引文
[1]王坚镪,吴琼,庄亦晖,等.耐碳青霉烯类肠杆菌科细菌的耐药机制探讨[J].检验医学,2017,32(12):1080-1084.
[2]Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing,28th ed. CLSI supplement M100-S28[S].Wayne PA:CLSI,2018:112-124.
[3]杨靖,时东彦,赵建宏,等.不同底物在CIM试验检测革兰阴性杆菌碳青霉烯酶中的应用评价[J].临床检验杂志,2016,34(8):575-578.
[4]张保荣,毕茹茹,顾兵.肠杆菌科细菌碳青霉烯酶表型检测方法研究进展[J].中国感染控制杂志,2018,17(2):175-179.
[5]Tamma PD,Opene BN,Gluck A,et al.Comparison of 11 phenotypic assays for accurate detection of carbapenemase-producing Enterobacteriaceae[J]. J Clin Microbiol,2017,55(4):1046-1055.
[6]Goodman KE,Simner PJ,Tamma PD,et al. Infection control implications of heterogeneous resistance mechanisms in carbapenemresistant Enterobacteriaceae(CRE)[J]. Expert Rev Infect Ther,2016,14(1):95-108.
[7]Teethaisong Y,Eumkeb G,Nakouti I,et al.A combined disc method with resazurin agar plate assay for early phenotypic screening of KPC,MBL and OXA-48 carbapenemases among Enterobacteriaceae[J]. J Appl Microbiol,2016,121(2):408-414.
[8]Yong D,Lee Y,Jeong SH,et al. Evaluation of double-disk potentiation and disk potentiation tests using dipicolinic acid for detection of metallo-β-lactamase-producing Pseudomonas spp. and Acinetobacter spp.[J]. J Clin Microbiol,2012,50(10):3227-3232.
[9]Giske CG,Gezelius L,Samuelsen O,et al. A sensitive and specific phenotypic assay for detection of metallo-β-lactamases and KPC in Klebsiella pneumoniae with the use of meropenem disks supplemented with aminophenylboronic acid,dipicolinic acid and cloxacillin[J].Clin Microbiol Infect,2011,17(4):552-556.
[10]Mac Vane SH,Crandon JL,Nichols WW,et al.In vivo efficacy of humanized exposures of ceftazidime-avibactam in comparison with ceftazidime against contemporary enterobacteriaceae isolates[J].Antimicrob Agents Chemother,2014,58(11):6913-6919.
[11]Drawz SM,Papp-Wallace KM,Bonomo RA.Newβ-lactamase inhibitors:a therapeutic renaissance in an MDR world[J]. Antimicrob Agents Chemother,2014,58(4):1835-1846.
[12]Yoshizumi A,Ishii Y,Livermore DM,et al. Efficacies of calciumEDTA in combination with imipenem in a murine model of sepsis caused by Escherichia coli with NDM-1β-lactamase[J]. J Infect Chemother,2013,19(5):992-995.
[2]Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing,28th ed. CLSI supplement M100-S28[S].Wayne PA:CLSI,2018:112-124.
[3]杨靖,时东彦,赵建宏,等.不同底物在CIM试验检测革兰阴性杆菌碳青霉烯酶中的应用评价[J].临床检验杂志,2016,34(8):575-578.
[4]张保荣,毕茹茹,顾兵.肠杆菌科细菌碳青霉烯酶表型检测方法研究进展[J].中国感染控制杂志,2018,17(2):175-179.
[5]Tamma PD,Opene BN,Gluck A,et al.Comparison of 11 phenotypic assays for accurate detection of carbapenemase-producing Enterobacteriaceae[J]. J Clin Microbiol,2017,55(4):1046-1055.
[6]Goodman KE,Simner PJ,Tamma PD,et al. Infection control implications of heterogeneous resistance mechanisms in carbapenemresistant Enterobacteriaceae(CRE)[J]. Expert Rev Infect Ther,2016,14(1):95-108.
[7]Teethaisong Y,Eumkeb G,Nakouti I,et al.A combined disc method with resazurin agar plate assay for early phenotypic screening of KPC,MBL and OXA-48 carbapenemases among Enterobacteriaceae[J]. J Appl Microbiol,2016,121(2):408-414.
[8]Yong D,Lee Y,Jeong SH,et al. Evaluation of double-disk potentiation and disk potentiation tests using dipicolinic acid for detection of metallo-β-lactamase-producing Pseudomonas spp. and Acinetobacter spp.[J]. J Clin Microbiol,2012,50(10):3227-3232.
[9]Giske CG,Gezelius L,Samuelsen O,et al. A sensitive and specific phenotypic assay for detection of metallo-β-lactamases and KPC in Klebsiella pneumoniae with the use of meropenem disks supplemented with aminophenylboronic acid,dipicolinic acid and cloxacillin[J].Clin Microbiol Infect,2011,17(4):552-556.
[10]Mac Vane SH,Crandon JL,Nichols WW,et al.In vivo efficacy of humanized exposures of ceftazidime-avibactam in comparison with ceftazidime against contemporary enterobacteriaceae isolates[J].Antimicrob Agents Chemother,2014,58(11):6913-6919.
[11]Drawz SM,Papp-Wallace KM,Bonomo RA.Newβ-lactamase inhibitors:a therapeutic renaissance in an MDR world[J]. Antimicrob Agents Chemother,2014,58(4):1835-1846.
[12]Yoshizumi A,Ishii Y,Livermore DM,et al. Efficacies of calciumEDTA in combination with imipenem in a murine model of sepsis caused by Escherichia coli with NDM-1β-lactamase[J]. J Infect Chemother,2013,19(5):992-995.