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肺炎克雷伯菌临床分离株产广谱和超广谱β-内酰胺酶的基因型研究

Study on the genotypes of spectrum β-lactamases and ESBLs in clinical isolates of Klebsiella pneumoniae

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【作者】 胡龙华余方友熊建球贾坤如胡晓彦况九龙桂炳东

【Author】 Hu Long-hua1,Yu Fang-you2,Xiong Jian-qiu1,Jia Kun-ru1,Hu Xiao-yan1,Kuang Jiu-long and Gui Bing-dong1(1 The Second Affiliated Hospital of Nanchang University,Nanchang 330006;2 The First Affiliated Hospital of Wenzhou Medical College,Wenzhou 325000)

【机构】 南昌大学第二附属医院温州医学院附属第一医院

【摘要】 目的了解产ESBLs肺炎克雷伯菌临床分离株的耐药性及其基因型分布特征,指导临床进行抗感染治疗,更好地控制耐药菌株的播散。方法对38株经双纸片试验确认为ESBLs表型阳性的肺炎克雷伯菌采用纸片扩散法进行药物敏感性试验;采用PCR法对ESBLs阳性株进行TEM型、SHV型、CTX-M型、VEB型和PER型ESBL基因检测,对PCR阳性产物进行测序确定基因型别;同时采用PCR法检测整合酶基因,对PCR产物进行测序确定整合酶类别。结果38株产ESBLs的肺炎克雷伯菌对青霉素类和头孢菌素类耐药性十分严重,对庆大霉素和复方磺胺甲唑的耐药率也均>90%,未检测到耐亚胺培南和美罗培南的菌株。38株ESBLs阳性菌株检测到3种ESBLs型,分别为TEM型、SHV型和CTX-M型,VEB型和PER型没有被检出。CTX-M型、TEM型和SHV型阳性率分别为92.1%(35/38)、84.2%(32/38)、76.3%(29/38)。整合酶基因阳性率为94.7%(36/38)。经测序,所有TEM型均为TEM-1广谱β内酰酶。29株SHV型阳性的PCR产物经测序证实,SHV-12有19株,SHV-11有4株,SHV-2有3株,SHV-28有2株及SHV-1有1株。35株CTX-M型中,CTX-M-14有22株,CTX-M-15有13株。同时检测到3种基因的有22株(57.9%),同时检测到2种基因的有14株(36.8%)。整合酶基因PCR产物经测序为Ⅰ类整合子。结论产ESBLs肺炎克雷伯菌临床分离株的耐药性及多重耐药性严重,Ⅰ类整合子是其多重耐药和耐药性传播的主要原因;CTX-M-14及SHV-12为肺炎克雷伯菌产ESBLs的主要基因型。

【Abstract】 Objective To investigate the genotypes of ESBLs in clinical isolates of Klebsiella pneumoniae isolated from Nanchang.Methods Thirty-eight isolates of ESBLs-producing Klebsiella pneumoniae,which were determined by double paper disk diffusion,were screened for antimicrobial susceptibility test.PCR detection of blaTEM,blaSHV,blaCTX-M,blaVEB,blaPER and intl was carried out with the previously reported oligonucleotide primers.The PCR products were sequenced on an ABI PRISM 3730 automated sequencer.Results The resistance of ESBLs-producing isolates to antimicrobial agents was serious.All tested isolates were susceptible to imipenem and meropenem.Three of 5 ESBL genotypes were detected.The positive rates of blaCTX-M、blaTEM and blaSHV were 92.1%(35/38),84.2%(32/38)and 76.3%(29/38),respectively.Thirty-six(94.7%)of 38 isolates were positive for intl.All TEM-type genes were TEM-1.Of 29 isolates harboring blaSHV,19,4,3 and 1 isolates carried blaSHV-12,blaSHV-11,blaSHV-1,and blaSHV-2,respectively.Twenty-two and 13 isolates harbored blaCTX-M-14 and blaCTX-M-15 Meanwhile 22(57.9%) and 14(36.8%) of 38 ESBLs-producing isolates carried three ESBLs genes and two ESBL genes.Conclusions blaCTX-M-14 and blaSHV-12 were main genotype of ESBLs genes in clinical isolates of ESBLs-producing Klebsiella pneumoniae.

【关键词】 肺炎克雷伯菌β-内酰胺酶基因型
【Key words】 Klebsiella pneumoniaeβ-lactamasesGenotype
  • 【文献出处】 中国抗生素杂志 ,Chinese Journal of Antibiotics , 编辑部邮箱 ,2009年06期
  • 【分类号】R450
  • 【被引频次】10
  • 【下载频次】100
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