耐碳青霉烯肺炎克雷伯菌耐药基因检测及其同源性分析
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摘要
目的:研究六安市人民医院耐碳青霉烯肺炎克雷伯菌(CRKP)的耐药基因及同源性。方法:收集2018年4-6月分离自临床标本的肺炎克雷伯菌40株,用法国梅里埃VITEK 2 COMPACT全自动细菌鉴定药敏仪进行鉴定和药敏。采用改良Hodge试验(MHT)和改良碳青霉烯灭活试验(mCIM)对40株菌进行碳青霉烯酶表型的筛选,采用PCR方法检测其是否携带已知的主要碳青霉烯耐药基因KPC、OXA-48、NDM-1。基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)对其进行同源性分析。结果:40株CRKP多分离自ICU病房(占55%),以呼吸道来源标本为主(占80%),对常用抗生素头孢菌素类、氨基糖苷类以及大环内酯类的耐药率均在80.0%以上。MHT和mCIM试验中有33株菌二者均为阳性,2株菌均为阴性,剩下5株MHT试验阳性而mCIM试验结果为阴性。32株菌携带KPC基因,未检出OXA-48及NDM-1基因。同源性分析结果显示40株CRKP分为7个型别,其中A型34株(占85%),B、C、D、E、F及G型各1株(各占2.5%)。结论:分离的CRKP以携带KPC-2基因为主,共有7个型别,以A型为主,为有效治疗和预防CRKP感染提供了参考依据。
Objective:To explore the drug resistance genes of carbapenem-resistant Klebsiella pneumonia(CRKP),and its homology analysis.Methods:Forty strains of Klebsiella pneumonia were isolated from clinical samples from April to June 2018,and the drug sensitivity of which was identified using VITEK 2 COMPACT automatic bacterial sensitivity instrument.The carbapenem phenotypes of the 40 strains of Klebsiella pneumonia were screened using the modified hodge test(MHT) and modified carbapenem inactivation test(mCIM).PCR method was used to detect the main carbapenem resistance genes(including KPC,OXA-48 and NDM-1 genes).Matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF MS) was used to analyze the homology of genes.Results:The 40 strains of CRKP were isolated from ICU ward(55%),mainly from respiratory tract samples(80%),and the resistance rates of those to cephalosporins,aminoglycosides and macrolides were above 80%.Thirty-three strains were positive in MHT test and mCIM test,2 strains were negative in MHT test and mCIM test,and the remaining 5 strains were positive in MHT test and negative in mCIM test.The KPC gene was detected in 32 strains,and the OXA-48 and NDM-1 genes were not detected.The results of homology analysis showed that 40 strains of CRKP were divided into 7 types,which included type A in 34 strains(85%),and type B,C,D,E,F and G in 1 strain,respectively(each accounting for 2.5%).Conclusions:The KPC-2 gene is the main genotype in CRKP,which can be divided into 7 kinds of types(type A as the main type).The study can provide the basis in effective treating and preventing the CRKP infection.
Objective:To explore the drug resistance genes of carbapenem-resistant Klebsiella pneumonia(CRKP),and its homology analysis.Methods:Forty strains of Klebsiella pneumonia were isolated from clinical samples from April to June 2018,and the drug sensitivity of which was identified using VITEK 2 COMPACT automatic bacterial sensitivity instrument.The carbapenem phenotypes of the 40 strains of Klebsiella pneumonia were screened using the modified hodge test(MHT) and modified carbapenem inactivation test(mCIM).PCR method was used to detect the main carbapenem resistance genes(including KPC,OXA-48 and NDM-1 genes).Matrix assisted laser desorption ionization time of flight mass spectrometry(MALDI-TOF MS) was used to analyze the homology of genes.Results:The 40 strains of CRKP were isolated from ICU ward(55%),mainly from respiratory tract samples(80%),and the resistance rates of those to cephalosporins,aminoglycosides and macrolides were above 80%.Thirty-three strains were positive in MHT test and mCIM test,2 strains were negative in MHT test and mCIM test,and the remaining 5 strains were positive in MHT test and negative in mCIM test.The KPC gene was detected in 32 strains,and the OXA-48 and NDM-1 genes were not detected.The results of homology analysis showed that 40 strains of CRKP were divided into 7 types,which included type A in 34 strains(85%),and type B,C,D,E,F and G in 1 strain,respectively(each accounting for 2.5%).Conclusions:The KPC-2 gene is the main genotype in CRKP,which can be divided into 7 kinds of types(type A as the main type).The study can provide the basis in effective treating and preventing the CRKP infection.
引文
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[19] 林琳,王晶,肖晓光,等.重症病房耐碳青霉烯肺炎克雷伯菌 KPC 基因检测及同源性分析[J].国际检验医学杂志,2018,39(7):798.
[20] 杨雪,刘琳,赵丹,等.耐碳青霉烯类肺炎克雷伯菌耐药基因检测与分子流行病学研究[J].重庆医学,2018(15):1977.
[21] 姚欣,冯莉,朱艮苗.碳青霉烯类耐药肺炎克雷伯菌分子流行病学及耐药机制[J].中国抗生素杂志,2018,43(1):85.
[22] FOURNIER PE,DRANCOURT M,COLSON P,et al.Modern clinical microbiology:new challenges and solutions[J].Nat Rev Microbiol,2013,11(8):574.
[23] SANGUINETTI M,POSTERARO B.Mass spectrometry applications in microbiology beyond microbe identification:progress and potential[J].Expert Eev Proteomics,2016,13(10):965.
[24] GIACOMETTI F,PIVA S,VRANCKX K,et al.Application of MALDI-TOF MS for the subtyping of Arcobacter butzleri strains and comparison with their MLST and PFGE types[J].Int J Food Microbiol,2018,277:50.
[25] LINDGREN ?,KARAMI N,KARLSSON R,et al.Development of a rapid MALDI-TOF MS based epidemiological screening method using MRSA as a model organism[J].Eur J Clin Microbiol Infect Dis,2018,37(1):57.
[2] 胡付品,朱德妹,汪复,等.2014年CHINET中国细菌耐药性监测[J].中国感染与化疗杂志,2015(5):401.
[3] 刘婧娴,俞静,李媛睿,等.肺炎克雷伯菌对碳青霉烯类抗生素的耐药机制研究[J].上海交通大学学报(医学版),2016,36(1):93.
[4] PATEL JB.Performance standards for antimicrobial susceptibility testing[M].Clin Laborat Stand Instit,2017.
[5] 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.
[6] 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.
[7] MARSH JW,KRAULAND MG,NELSON JS,et al.Genomic epidemiology of an endoscope-associated outbreak of Klebsiella pneumoniae carbapenemase (KPC)-producing K.pneumoniae[J].PLoS One,2015,10(12):e0144310.
[8] YIGIT H,QUEENAN AM,RASHEED JK,et al.Carbapenem-resistant strain of Klebsiella oxytoca harboring carbapenem-hydrolyzing β-lactamase KPC-2[J].Antimicrob Agents Chemother,2003,47(12):3881.
[9] ENDIMIANI A,CHOUDHARY Y,BONOMO RA.In vitro activity of NXL104 in combination with β-lactams against Klebsiella pneumoniae isolates producing KPC carbapenemases[J].Antimicrob Agents Chemother,2009,53(8):3599.
[10] CAMPOS AC,ALBIERO J,ECKER AB,et al.Outbreak of Klebsiella pneumoniae carbapenemase-producing K pneumoniae:a systematic review[J].Am J Infect Control,2016,44(11):1374.
[11] OKAMOTO K,LIN MY,HAVERKATE M,et al.Modifiable risk factors for the spread of Klebsiella pneumoniae carbapenemase-producing Enterobacteriaceae among long-term acute-care hospital patients[J].Infect Control Hosp Epidemiol,2017,38(6):670.
[12] GALLAGHER JC,KURIAKOSE S,HAYNES K,et al.A case-case-control study of patients with carbapenem-resistant and third-generation cephalosporin-resistant Klebsiella pneumoniae bloodstream infections[J].Antimicrob Agents Chemother,2014,58(10):5732.
[13] 王凤,赖开生,安静娜,等.2013-2017 年某三甲医院产 KPC 肺炎克雷伯菌流行特征研究[J].中华医院感染学杂志,2017,27(24):5521.
[14] 赵金云,许文芳,金法祥.ICU 和非 ICU 患者感染肺炎克雷伯菌中超广谱 β-内酰胺酶和碳青霉烯酶检测[J].中华医院感染学杂志,2018,28(2):165.
[15] 凌勇,肖洪广,邱芳华.耐碳青霉烯肺炎克雷伯菌的耐药基因分析[J].热带医学杂志,2018,18(3):358.
[16] PORRECA AM,SULLIVAN KV,GALLAGHER JC.The Epidemiology,Evolution,and Treatment of KPC-Producing Organisms[J].Curr Infect Dis Rep,2018,20(6):13.
[17] 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,2016,32(23):3208.
[18] 高倩倩,殷杏,祝俊英,等.碳青霉烯类耐药肺炎克雷伯菌的分子特征[J].中国感染与化疗杂志,2013,21(9):52.
[19] 林琳,王晶,肖晓光,等.重症病房耐碳青霉烯肺炎克雷伯菌 KPC 基因检测及同源性分析[J].国际检验医学杂志,2018,39(7):798.
[20] 杨雪,刘琳,赵丹,等.耐碳青霉烯类肺炎克雷伯菌耐药基因检测与分子流行病学研究[J].重庆医学,2018(15):1977.
[21] 姚欣,冯莉,朱艮苗.碳青霉烯类耐药肺炎克雷伯菌分子流行病学及耐药机制[J].中国抗生素杂志,2018,43(1):85.
[22] FOURNIER PE,DRANCOURT M,COLSON P,et al.Modern clinical microbiology:new challenges and solutions[J].Nat Rev Microbiol,2013,11(8):574.
[23] SANGUINETTI M,POSTERARO B.Mass spectrometry applications in microbiology beyond microbe identification:progress and potential[J].Expert Eev Proteomics,2016,13(10):965.
[24] GIACOMETTI F,PIVA S,VRANCKX K,et al.Application of MALDI-TOF MS for the subtyping of Arcobacter butzleri strains and comparison with their MLST and PFGE types[J].Int J Food Microbiol,2018,277:50.
[25] LINDGREN ?,KARAMI N,KARLSSON R,et al.Development of a rapid MALDI-TOF MS based epidemiological screening method using MRSA as a model organism[J].Eur J Clin Microbiol Infect Dis,2018,37(1):57.