【摘要】 目的探讨碳青霉烯类耐药肺炎克雷伯菌产β-内酰胺酶情况和β-内酰胺酶基因分型研究。方法用Vitek-Ⅱ全自动微生物分析仪对本院临床标本中分离的对碳青霉烯类耐药的肺炎克雷伯菌进行菌种鉴定和药物敏感试验;用冻融法提取β-内酰胺酶,三维试验检测β-内酰胺酶[AmpC酶、超广谱β-内酰胺酶(ESBLs)、金属酶];改良Hodge试验检测KPC酶;用PCR扩增TEM、SHV、CTX-M、PER、VEB、DHA、MIR/ACT、KPC、IMP、VIM、SPM、GIM和NDM-1基因,并对阳性基因进行测序分析确定其基因型;REP-PCR检测分析其同源性。结果 4株对碳青霉烯类抗生素耐药肺炎克雷伯菌均表现为多重耐药,其中2株菌对所有测试的抗菌药物均耐药;1株菌产金属酶,由IMP-4型金属酶基因编码;4株菌均产生DHA型AmpC酶,2株菌产CTX-M-14型ESBLs;1株菌产TEM-71型ESBLs,均经测序证实;未检测出SHV、PER、VEB、MIR/ACT、KPC、VIM、SPM、GIM和NDM-1基因型。REP-PCR显示4株菌属于3个不同的克隆型。结论本院出现碳青霉烯类耐药的肺炎克雷伯菌,属于不同克隆型,产多种β-内酰胺酶(AmpC酶、ESBLs、金属酶),基因型为DHA、IMP-4、CTX-M-14、TEM-71。更多还原

【Abstract】 Objective To detect the β-lactamases and genetyping of carbapenem-resistant Klebsiella pneumoniae.Methods The strains and antibiotic susceptibility were identified by VITEK-Ⅱ.β-lactamases(AmpC,ESBLs,MBLand KPC) were screened by three dimensional test and improved Hodge test.REP-PCR was performed to analyze homology.Specific PCR for detection of TEM,SHV,CTX-M,PER,VEB,DHA,MIR/ACT,KPC,IMP,VIM,SPM,GIM,NDM-1 and DNA sequencing were performed to confirm genotypes.The homology of carbapenem-resistant Klebsiella pneumoniae was detect by REP-PCR.Results Four Klebsiella pneumoniae isolates were screened to be resistant to most antimicrobials.Two strains were resistant to all the antimicrobials tested.REP-PCR showed that the four isolates belong to three different clonal strains.Four strains produced DHA AmpC,1 strain produced IMP-4 metallo-β-lactamase and 3 strains produced ESBLs(CTX-M-14 and TEM-71).Specific PCR and DNA sequence analysis indicated that the strains were mediated by DHA,IMP-4,CTX-M-14,TEM-71 genes.SHV,PER,VEB,MIR/ACT,KPC,VIM,SPM,GIM and NDM-1 genes were undetectable.Conclusion The Klebsiella pneumoniae isolates with carbapenem resistance obtained from our hospital belonged to different clones which produced metallo-β-lactamase,AmpC and ESBLs.The gene types were IMP-4,DHA,CTX-M-14 and TEM-71.更多还原