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目的对医院感染常见的革兰阴性杆菌产超广谱β-内酰胺酶(ESBLs)、AmpC酶耐药性进行分析,为临床合理用药提供依据。方法收集住院部各科室临床细菌样本,用VITEK 2 Compact系统进行鉴定和药敏试验。采用药敏纸片法筛选产ESBLs、AmpC酶菌株。采用PCR通用引物扩增AmpC酶与ESBLs基因并进行序列测定以确定其基因亚型。结果 66株革兰阴性杆菌中产ESBLs酶的细菌以大肠杆菌为主(72.1%),其次为肺炎克雷伯菌(64.7%),产AmpC酶以铜绿假单胞菌及阴沟肠杆菌为主(均为100.0%),同时产ESBLs+AmpC酶以铜绿假单胞菌为主(33.3%)。检出4种ESBLs酶基因(TEM、SHV、OXA和CTX-M)、3种AmpC酶基因(DHA、CIT、EBC)。ESBLs中以CTX-M基因型检出率最高(62.1%);AmpC中以DHA基因型检出率最高(18.2%)。结论本院存在同时产ESBLs、AmpC酶革兰阴性杆菌流行株,上述临床常见病原菌携带多类药物钝化酶基因并存在不同的优势基因携带模式。
Objective To analyze the antibiotic resistance of extended-spectrum β-lactamase (ESBLs) and AmpC enzyme in common Gram-negative bacilli of nosocomial infections and provide basis for clinical rational drug use. Methods Clinical bacterial samples collected from departments of inpatient department were collected and identified by VITEK 2 Compact system and susceptibility testing. Drug susceptibility screening method using ESBLs, AmpC enzyme strains. AmpC gene and ESBLs gene were amplified by PCR universal primers and sequenced to determine their gene subtypes. Results Among the 66 Gram-negative bacilli, ESBLs-producing bacteria were Escherichia coli (72.1%), Klebsiella pneumoniae (64.7%), AmpCases were Pseudomonas aeruginosa and Enterobacter cloacae Both 100.0%), while producing ESBLs + AmpC enzyme Pseudomonas aeruginosa (33.3%). Four kinds of ESBLs genes (TEM, SHV, OXA and CTX-M) and three kinds of AmpC enzyme genes (DHA, CIT, EBC) were detected. The highest detection rate of CTX-M genotypes was found in ESBLs (62.1%). The highest detection rate of DHA genotypes was in AmpC (18.2%). Conclusion There are ESBLs-producing and AmpC-Gram-negative bacilli strains in our hospital. The common clinical pathogens carry multiple types of drug-inactivating enzyme genes and different dominant gene carrying patterns exist.