目的 分析本院临床分离标本中超广谱β-内酰胺酶(ESBLs)大肠埃希菌和肺炎克雷伯菌的耐药性及相关耐药基因的检出情况.方法 采用纸片扩散法(K-B法)对从我院分离的236株大肠埃希菌和224株肺炎克雷伯菌进行ESBLs表型鉴定、药敏试验,利用PCR法扩增ESBLs耐药相关基因TEM、SHV和CTX-M(M1、M2和M9),对其产物进行测序分析验证.结果 产ESBLs大肠埃希菌的检出率为32.63%,产ESBLs肺炎克雷伯菌的检出率为30.36%;ESBLs大肠埃希菌、ESBLs肺炎克雷伯菌对氨苄西林、哌拉西林、头孢呋辛、头孢他啶、 头孢曲松、 氨曲南、 庆大霉素、 左氧氟沙星等的耐药率均高于相应的非产ESBLs菌(P均<0.05);产ESBLs肺炎克雷伯菌SHV基因、CTX-M2亚型检出率均高于产ESBLs大肠埃希菌(P<0.05),CTX-M9亚型的检出率低于产ESBLs大肠埃希菌(P<0.05);所有产物测序分析结果与PCR扩增结果一致.结论 产ESBLs的大肠埃希菌和肺炎克雷伯菌耐药形势复杂、多样,临床应重视常规药敏试验并结合耐药基因检测的应用.%Objective To investigate the antibiotic resistance and genotype in ESBLs Escherichia coli and Klebsiella pneumoniae.Methods 236 Escherichia coli and 224 Klebsiella pneumoniae strains were continuously collected from our hospital and they were tested by antibiotic sensitivity and phonotype of ESBLs. ESBLs related genes of TEM,SHV and CTX-M(M1,M2 and M9)were amplified by PCR and verified by sanger sequencing.Results The positive rate of ESBLs Escherichia coli was 32.63% and that of Klebsiella pneumoniaeis was 30.36%. Both ESBLs Escherichia coli and ESBLs Klebsiella pneumoniae were highly resistant to Ampicillin,Piperacillin,Cefuroxime,Ceftazidime,Ceftriaxone,Aztreonam,Gentamycin,Levofloxacin,etc. As for resistant genes detecting results,the SHV and CTX-M2 gene in ESBLs Escherichia colistrains was significantly higher than that in ESBLs Escherichia coli with significant difference(P<0.05)while CTX-M9 gene in ESBLs Escherichia colistrains was significantly lower than that in ESBLs Escherichia coli.Conclusion The situation of the antibiotic-resistance of ESBLs Escherichia coli and Klebsiella pneumoniaeare is serious in our hospital and it must be attached more attention to the combination with routine test and antibiotic-resistant gene detection which are applied in clinics.
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