改良Hodge与Carba NP和mCIM试验快速检测产碳青霉烯酶肺炎克雷伯菌的方法学比较
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摘要
目的比较改良Hodge试验、Carba NP试验及碳青霉烯灭活试验(mCIM试验)对产碳青霉烯酶肺炎克雷伯菌快速表型筛选的能力。方法以PCR扩增肺炎克雷伯菌中携带的碳青霉烯酶相关耐药基因为金标准,比较改良Hodge试验、Carba NP试验及mCIM试验对202株碳青霉烯酶肺炎克雷伯菌试验菌的筛选能力。结果 202株试验菌经PCR扩增120株耐药基因阳性,产物测序得知75株只携带bla_(KPC-2)基因,20株同时携带bla_(KPC-2)和bla_(VIM-2)基因,19株同时携带bla_(KPC-2)和bla_(IMP-4)基因,3株只携带blaIMP-4基因,2株只携带bla_(VIM-2)基因,1株只携带bla_(NDM-1)基因;120株产碳青霉烯酶试验组菌株对亚胺培南、厄他培南及头孢菌素类抗菌药物的耐药率均为100.00%,对替加环素和多黏菌素B敏感。82株非产碳青霉烯酶的对照菌株对亚胺培南、厄他培南均敏感,对头孢曲松的耐药率为100.00%。以PCR结果为金标准,试验菌通过改良Hodge试验检测的灵敏度为92.50%,特异性为97.56%;Carba NP试验检测的灵敏度为94.17%,特异性为98.78%;mCIM试验检测的灵敏度为98.33%,特异性为100.00%。与PCR结果相比,改良Hodge试验的一致率为94.55%,Kappa值为0.8885;Carba NP试验的一致率为96.04%,Kappa值为0.9188;mCIM试验的一致率为99.01%,Kappa值为0.9796。结论 Carba NP试验和mCIM试验在快速检测产碳青霉烯酶的肺炎克雷伯菌较改良Hodge试验具有更高的敏感性和特异性,并且操作简便,适合在临床微生物学实验室、医院感染控制室及疾病预防控制系统各单位普及应用。
OBJECTIVE To compare the capability of modified Hodge test,modified carbapenem inactivation method(mCIM)and Carba NP test for rapid screening of phenotypes of carbapenemase-producing Klebsiella pneumoniae.METHODS PCR was used to amplify the carbapenemase-related genes in the K.pneumoniae and taken as the golden standard,and the capability of the modified Hodge test,Carba NP test and mCIM test in screening of 202 strains of carbapenemase-producing K.pneumoniae test isolates was observed and compared.RESULTS Of the 202 test strains,120 were PCR amplified positive for the drug resistance genes,the result of sequencing of amplified products showed that 75 strains only carried bla_(KPC) gene,20 strains simultaneously carried bla_(KPC) and bla_(VIM) genes,19 strains simultaneously carried bla_(KPC) and bla_(IMP) genes,3 strains only carried bla_(IMP) gene,2 strains only carried blaVIM gene,and 1 strain only carried blaNDM-1 gene.The drug resistance rates of the 120 strains of carbapenemase-producing K.pneumoniae isolates to imipenem,ertapenem and cephalosporins were 100%,while the strains were sensitive to tigecycline and polymyxin B.The 82 strains of non-carbapenemase-producing control isolates were sensitive to imipenem and ertapenem,while the drug resistance rate to ceftriaxone was 100%.Taken the PCR result as the golden standard,the sensitivities of the modified Hodge test,Carba NP test and mCIM test in screening of the test strains were 92.5%,94.17% and 98.33%,respectively;the specificities were 97.56%,98.78% and 100%,respectively.As compared with the PCR results,the consistency rate of the modified Hodge test was 94.55%,with the Kappa value 0.8885;the consistency rate of the Carba NP test was 96.04%,with Kappa value 0.9188;the consistency rate of the mCIM test was 99.01%,with Kappa value 0.9796.CONCLUSIONThe sensitivity and specificity of the Carba NP test and mCIM test are higher than those of the modified Hodge test in rapid detection of carbapenemase-producing K.pneumoniae,and the operation procedures are so simple that they are worthy to be promoted in clinical microorganism laboratory,infection control rooms and departments of disease prevention and control.
OBJECTIVE To compare the capability of modified Hodge test,modified carbapenem inactivation method(mCIM)and Carba NP test for rapid screening of phenotypes of carbapenemase-producing Klebsiella pneumoniae.METHODS PCR was used to amplify the carbapenemase-related genes in the K.pneumoniae and taken as the golden standard,and the capability of the modified Hodge test,Carba NP test and mCIM test in screening of 202 strains of carbapenemase-producing K.pneumoniae test isolates was observed and compared.RESULTS Of the 202 test strains,120 were PCR amplified positive for the drug resistance genes,the result of sequencing of amplified products showed that 75 strains only carried bla_(KPC) gene,20 strains simultaneously carried bla_(KPC) and bla_(VIM) genes,19 strains simultaneously carried bla_(KPC) and bla_(IMP) genes,3 strains only carried bla_(IMP) gene,2 strains only carried blaVIM gene,and 1 strain only carried blaNDM-1 gene.The drug resistance rates of the 120 strains of carbapenemase-producing K.pneumoniae isolates to imipenem,ertapenem and cephalosporins were 100%,while the strains were sensitive to tigecycline and polymyxin B.The 82 strains of non-carbapenemase-producing control isolates were sensitive to imipenem and ertapenem,while the drug resistance rate to ceftriaxone was 100%.Taken the PCR result as the golden standard,the sensitivities of the modified Hodge test,Carba NP test and mCIM test in screening of the test strains were 92.5%,94.17% and 98.33%,respectively;the specificities were 97.56%,98.78% and 100%,respectively.As compared with the PCR results,the consistency rate of the modified Hodge test was 94.55%,with the Kappa value 0.8885;the consistency rate of the Carba NP test was 96.04%,with Kappa value 0.9188;the consistency rate of the mCIM test was 99.01%,with Kappa value 0.9796.CONCLUSIONThe sensitivity and specificity of the Carba NP test and mCIM test are higher than those of the modified Hodge test in rapid detection of carbapenemase-producing K.pneumoniae,and the operation procedures are so simple that they are worthy to be promoted in clinical microorganism laboratory,infection control rooms and departments of disease prevention and control.
引文
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[14]Cuzon G,Bentchouala C,Vogel A,et al.First outbreak of OXA-48-positive carbapenem-resistant Klebsiella pneumoniae isolates in Constantine,Algeria[J].Int J Antimicrob Agents,2015,46(6):725-727.
[15]Jamal WY,Albert MJ,Rotimi VO.High prevalence of New Delhi metallo-β-lactamase-1(NDM-1)producers among carbapenem-resistant Enterobacteriaceae in Kuwait[J].PLoSOne,2016,11(3):e0152638.
[16]Stillwell T,Green M,Barbadora K,et al.Outbreak of KPC-3producing carbapenem-resistant Klebsiella pneumoniae in a US pediatric hospital[J].J Pediatric Infect Dis Soc,2015,4(4):330-338.
[17]Torres-Gonzalez P,Bobadilla-Del Valle M,Tovar-Calderon E,et al.Outbreak caused by Enterobacteriaceae harboring NDM-1metallo-β-lactamase carried in an IncFII plasmid in a tertiary care hospital in Mexico City[J].Antimicrob Agents Chemother,2015,59(11):7080-7083.
[18]Munoz-Price LS,Poirel L,Bonomo RA,et al.Clinical epidemiology of the global expansion of Klebsiella pneumoniae carbapenemases[J].Lancet Infect Dis,2013,13(9):785-796.
[19]胡仁静,严子禾,胡锡池.Carba NP试验及CIM试验检测肠杆菌科细菌碳青霉烯酶的性能评估[J].职业与健康,2016(23):3208-3213.
[20]Nordmann P,Poirel L.Strategies for identification of carbapenemase-producing Enterobacteriaceae[J].J Antimicrob Chemother,2013,68(3):487.
[21]朱雄,袁敏,刘佳,等.CarbaNP碳青霉烯酶检测方法在医院感染监测中的应用[J].中华医院感染学杂志,2014,24(23):5721-5724.
[22]Bayramoglu G,Ulucam G,Gencoglu Ozgur C.Evaluation of carbapenem inactivation method for the identification of carbapenemase-producing Enterobacteriaceae strains[J].Mikrobiyol Bul,2016,50(3):505-507.
[2]Du J,Li P,Liu H,et al.Phenotypic and molecular characterization of multidrug-resistant Klebsiella pneumoniae isolated from a university teaching hospital,China[J].PLoS One,2014,9(4):e95181.
[3]陈金云,傅鹰,杨青,等.KPC-2及IMP-4酶介导肠杆菌科细菌碳青霉烯类耐药研究[J].中华微生物学和免疫学杂志,2015(6):419-426.
[4]赵燕,袁轶群,叶琴,等.耐碳青霉烯类肺炎克雷伯菌碳青霉烯酶基因检测[J].中华医院感染学杂志,2013,23(17):4081-4083.
[5]马维佳,徐绣宇,孙坤玲,等.Carba NP法与改良Hodge试验检测产碳青霉烯酶肠杆菌的方法学评价[J].中华微生物学和免疫学杂志,2016,36(1):64-65.
[6]Clinical and Laboratory Standards Institute.Performance standards for antimicrobial suscetibility testing:twenty-second informational supplement.CLSI document M100-S25[S].Wayne,Pa:CLSI,2015.
[7]张雅薇,王辉.2015年CLSI M100-S25主要更新内容介绍[J].中华检验医学杂志,2015,38(4):229-232.
[8]Clinical and Laboratory Standards Institute.Performance standards for antimicrobial suscetibility testing:twenty-second informational supplement.CLSI document M100-S27[S].Wayne,Pa:CLSI,2017.
[9]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 assessphenotypic carbapenemase activity in gram negative rods[J].PLoS One,2015,10(3):e0123690.
[10]Hong SS,Kyeongmi K,Young HJ,et al.Multiplex PCR for rapid detection of genes encoding class A carbapenemases[J].Ann Lab Med,2012,32(5):359-361.
[11]Yan J,Hsueh P,Ko W,et al.Metallo-beta-lactamases in clinical Pseudomonas isolates in Taiwan and identification of VIM-3,a novel variant of the VIM-2enzyme[J].Antimicrob Agents Chemother,2001,45(8):2224-2228.
[12]Poirel L,Walsh TR,Cuvillier V,et al.Multiplex PCR for detection of acquired carbapenemase genes[J].Diagn Microbiol Infect Dis,2011,70(1):119-123.
[13]Yigit H,Queenan AM,Anderson GJ,et al.Novel carbapenem-hydrolyzing beta-lactamase,KPC-1,from a carbapenem-resistant strain of Klebsiella pneumoniae[J].Antimicrob A-gents Chemother,2001,45(4):1151-1161.
[14]Cuzon G,Bentchouala C,Vogel A,et al.First outbreak of OXA-48-positive carbapenem-resistant Klebsiella pneumoniae isolates in Constantine,Algeria[J].Int J Antimicrob Agents,2015,46(6):725-727.
[15]Jamal WY,Albert MJ,Rotimi VO.High prevalence of New Delhi metallo-β-lactamase-1(NDM-1)producers among carbapenem-resistant Enterobacteriaceae in Kuwait[J].PLoSOne,2016,11(3):e0152638.
[16]Stillwell T,Green M,Barbadora K,et al.Outbreak of KPC-3producing carbapenem-resistant Klebsiella pneumoniae in a US pediatric hospital[J].J Pediatric Infect Dis Soc,2015,4(4):330-338.
[17]Torres-Gonzalez P,Bobadilla-Del Valle M,Tovar-Calderon E,et al.Outbreak caused by Enterobacteriaceae harboring NDM-1metallo-β-lactamase carried in an IncFII plasmid in a tertiary care hospital in Mexico City[J].Antimicrob Agents Chemother,2015,59(11):7080-7083.
[18]Munoz-Price LS,Poirel L,Bonomo RA,et al.Clinical epidemiology of the global expansion of Klebsiella pneumoniae carbapenemases[J].Lancet Infect Dis,2013,13(9):785-796.
[19]胡仁静,严子禾,胡锡池.Carba NP试验及CIM试验检测肠杆菌科细菌碳青霉烯酶的性能评估[J].职业与健康,2016(23):3208-3213.
[20]Nordmann P,Poirel L.Strategies for identification of carbapenemase-producing Enterobacteriaceae[J].J Antimicrob Chemother,2013,68(3):487.
[21]朱雄,袁敏,刘佳,等.CarbaNP碳青霉烯酶检测方法在医院感染监测中的应用[J].中华医院感染学杂志,2014,24(23):5721-5724.
[22]Bayramoglu G,Ulucam G,Gencoglu Ozgur C.Evaluation of carbapenem inactivation method for the identification of carbapenemase-producing Enterobacteriaceae strains[J].Mikrobiyol Bul,2016,50(3):505-507.