耐碳青霉烯类肺炎克雷伯菌临床分离株中喹诺酮类及16SrRNA甲基化酶基因的检测
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摘要
目的了解临床分离株中耐碳青霉烯肺炎克雷伯菌(CRKP)的氟喹诺酮类耐药基因及16SrRNA甲基化酶基因存在情况和药敏特点。方法收集临床标本中分离的6株CRKP,采用VITEK 2compact全自动微生物鉴定和药敏系统鉴定细菌及药敏试验,采用聚合酶链反应(PCR)扩增氟喹诺酮类耐药基因和16SrRNA甲基化酶基因。结果 6株CRKP临床分离株呈现多药耐药特点,对环丙沙星和左氧氟沙星的耐药率分别是83.3%和83.3%,对庆大霉素、阿米卡星和妥布霉素的耐药率分别是66.7%,50%和66.7%。检出喹诺酮类耐药相关基因qnrB,qnrS和qnrD,基因阳性率分别为50%,50%和33.3%,未检出qnrA,qnrC和qepA基因。检出16SrRNA甲基化酶耐药相关基因armA,基因阳性率为50%,未检出rmtA,rmtB,rmtC和npmA基因。结论 6株CRKP对喹诺酮类和氨基糖类抗生素耐药严重,其耐药性与喹诺酮类耐药基因及16SrRNA甲基化酶基因高度相关,其耐药基因主要型别分别是qnrB,qnrS,qnrD和armA。
Objective To understand the presence of quinolone resistance gene and 16 SrRNA methylation enzyme gene and the characteristics of drug sensitivity in the clinical isolated carbapenem-resistant Klebsiella pneumoniae(CRKP).Methods 6 strains of CRKP isolated from clinical specimens were collected,and bacteria and drug sensitivity tests were identified by VITEK 2 compact automatic microbial identification and drug sensitivity system,and polymerase chain reaction(PCR)was used to amplify the quinolone resistance gene and 16 SrRNA methylase gene.Results 6 strains of CRKP clinical isolates showed multidrug resistance,and the resistance rates for ciprofloxacin and levofloxacin were 83.3%and 83.3%respectively.The resistance rates for gentamicin,amikacin and tobramycin were 66.7%,50% and 66.7%,respectively.The positive rates of qnrB,qnrS and qnrD genes were 50%,50% and 33.3%,respectively.The negative rates of qnrA,qnrC and qepA were found.The 16 SrRNA methylase resistance related genes were detected.The positive rate of armA gene was 50%,and rmtA,rmtB,rmtC and npmA genes were negative.Conclusion 6 strains of CRKP were resistant to quinolones and aminoglycos,and their resistance was highly correlated with quinolone resistance genes and 16 SrRNA methylase genes.The main types of drug resistance genes were qnrB,qnrS,qnrD and armA.
Objective To understand the presence of quinolone resistance gene and 16 SrRNA methylation enzyme gene and the characteristics of drug sensitivity in the clinical isolated carbapenem-resistant Klebsiella pneumoniae(CRKP).Methods 6 strains of CRKP isolated from clinical specimens were collected,and bacteria and drug sensitivity tests were identified by VITEK 2 compact automatic microbial identification and drug sensitivity system,and polymerase chain reaction(PCR)was used to amplify the quinolone resistance gene and 16 SrRNA methylase gene.Results 6 strains of CRKP clinical isolates showed multidrug resistance,and the resistance rates for ciprofloxacin and levofloxacin were 83.3%and 83.3%respectively.The resistance rates for gentamicin,amikacin and tobramycin were 66.7%,50% and 66.7%,respectively.The positive rates of qnrB,qnrS and qnrD genes were 50%,50% and 33.3%,respectively.The negative rates of qnrA,qnrC and qepA were found.The 16 SrRNA methylase resistance related genes were detected.The positive rate of armA gene was 50%,and rmtA,rmtB,rmtC and npmA genes were negative.Conclusion 6 strains of CRKP were resistant to quinolones and aminoglycos,and their resistance was highly correlated with quinolone resistance genes and 16 SrRNA methylase genes.The main types of drug resistance genes were qnrB,qnrS,qnrD and armA.
引文
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[9]李娟,李从容,黄俊,等.肺炎克雷伯菌中质粒介导耐氟喹诺酮类耐药基因qnr的流行现状及耐药特征[J].现代检验医学杂志,2008,23(4):28-29.Li J,Li CR,Huang J,et al.Prevalence of qnr gene of plasmid-mediated quionlone resistance in Klebsiella pneunoniae[J].J Mod Lab Med,2008,23(4):28-29.
[10] Yu F,Wang L,Pan J,et al.Prevalence of 16SrRNA methylase genes in Klebsiella pneumoniae isolates from a Chinese teaching hospital:coexistence of rmtB and armA genes in the same isolate[J].Diagn Microbial Infect Dis,2009,64(1):57-63.
[11]杭亚平,宁长秀,汪红,等.耐碳青霉烯类抗生素肺炎克雷伯菌临床分离株中16SrRNA甲基化酶基因的检测[J].中国临床药理学杂志,2014,30(4):327-331.Hang YP,Ning CX,Wang H,et al.Detection of 16S rRNA methylase genes in clinical isolates carbapenem-resistant Klebsiella-pneumoniae[J].Chin J Clin Pharmacol,2014,30(4):327-331.
[2] Cattoir V,Poirel L,Rotimi V,et al.Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing enterobacterial isolates[J].J Antimicrob Chemother,2007,60(2):394-397.
[3] Kim HB,Park CH,Kim CJ,et al.Prevalence of plasmid-mediated quinolone resistance determinants over a 9-year period[J].Antimicrob Agents Chemother,2009,53(2):639-645.
[4] Liu JH,Deng YT,Zeng ZL,et al.Coprevalence of plasmid-mediated quinolone resistance determinants QepA,Qnr,and AAC(6’)-Ib-cr among 16SrRNA methylase RmtB-producing Escherichia coli isolates from pigs[J].Antimicrob Agents Chemother,2008,52(8):2992-2993.
[5] Zhao JY,Dang H.Coastal seawater bacteria harbor a large reservoir of plasmid-mediated quinolone resistance determinants in Jiaozhou Bay,China[J].Microb Ecol,2012,64(1):187-199.
[6] Bercot B,Poirel L,Nordmann P.Updated multiplex polymerase chain reaction for detection of 16SrRNA methylases:high prevalence among NDM-1producers[J].Diagn Microbiol Infect Dis,2011,71(4):442-445.
[7]张丽,朱元祺,张小兵,等.耐碳青霉烯类肺炎克雷伯菌的耐药基因检测[J].中华医院感染学杂志,2014,24(23):5734-5736,5745.Zhang L,Zhu YQ,Zhang XB,et al.Detection of genotypes of carbapenem-resistant Klebsiella pneumoniae isolates[J].Chinese Journal of Nosocomiology,2014,24(23):5734-5736,5745.
[8] Ma JY,Zeng ZL,Chen ZL,et al.High prevalence of plasmid-mediated quinolone resistance determinants qnr,aac(6’)-Ib-cr,and qepA among ceftiofur-resistant Enterobacteriaceae isolates from companion and food-producing animals[J].Antimicrob Agents Chemother,2009,53(2):519-524.
[9]李娟,李从容,黄俊,等.肺炎克雷伯菌中质粒介导耐氟喹诺酮类耐药基因qnr的流行现状及耐药特征[J].现代检验医学杂志,2008,23(4):28-29.Li J,Li CR,Huang J,et al.Prevalence of qnr gene of plasmid-mediated quionlone resistance in Klebsiella pneunoniae[J].J Mod Lab Med,2008,23(4):28-29.
[10] Yu F,Wang L,Pan J,et al.Prevalence of 16SrRNA methylase genes in Klebsiella pneumoniae isolates from a Chinese teaching hospital:coexistence of rmtB and armA genes in the same isolate[J].Diagn Microbial Infect Dis,2009,64(1):57-63.
[11]杭亚平,宁长秀,汪红,等.耐碳青霉烯类抗生素肺炎克雷伯菌临床分离株中16SrRNA甲基化酶基因的检测[J].中国临床药理学杂志,2014,30(4):327-331.Hang YP,Ning CX,Wang H,et al.Detection of 16S rRNA methylase genes in clinical isolates carbapenem-resistant Klebsiella-pneumoniae[J].Chin J Clin Pharmacol,2014,30(4):327-331.