摘要
目的分离与鉴定海南地区临床发现的耐碳青酶烯类药物的产气肠杆菌是否携带新德里金属β-内酰胺酶-1(New Delhi metallo-beta-lactamas-1,NDM-1)基因。方法用法国梅里埃API 20E生化鉴定条和VITEK2全自动细菌鉴定仪及其配套CNS试剂,鉴定出临床耐亚胺培南和美洛培南的产气肠杆菌,并用亚胺培南/亚胺培南+EDTA纸片法协同试验测定产金属酶类碳青霉烯酶;然后,对目的菌株进行NDM-1耐药基因聚合酶链反应(polymerase chain reaction,PCR)特异性扩增后,将扩增产物行琼脂糖凝胶电泳和测序鉴定及BLAST比对,以确定其NDM-1基因。结果采用亚胺培南/亚胺培南+EDTA纸片法协同试验,从10株耐亚胺培南或美洛培南的产气肠杆菌中,检出4株金属类碳青霉烯酶表型阳性菌株。对其中4株金属酶表型阳性的产气肠杆菌进行NDM-1基因PCR扩增后,对其产物进行琼脂糖凝胶电泳和测序分析,确认1株为携带NDM-1的耐药基因菌株。结论海南发现1株携带bla NDM-1基因的产气肠杆菌。
Objective To isolate and confirm the reduced carbapenem susceptible Enterobacter aerogene with NDM-1 （New Delhi metallo-beta-lactamas-1） in Hainan. Methods The positive Enterobacter aerogene strains, which had the ability to hydrolyzing Imiperem and Meropenem, were discerned by BioMerieux API 20E identification system and automatic VITEK-2 dentification system. The production of metalloprotease carbapenemase was measured synergically using imipenem/imipenem＋EDTA paper method. The specific NDM-1 genes was identified by sequencing of products by polymerase chain reaction（PCR） amplification. Results From ten to four positive strains were identified as metalloprotease carbapenemase positive in 10 imipenem or meropenem- resistant Enterobacter aerogene by imipenem/imipenem＋EDTA paper method. One strain was confirmed as NDM- 1 positive strain by PCR sequencing. Conclusion One Enterobacter aerogene strain carrying blaNDM-1 gene was identified in Hainan.
出处
《中华临床实验室管理电子杂志》
2015年第2期109-112,共4页
Chinese Journal of Clinical Laboratory Management(Electronic Edition）
基金
海南省自然基金资助项目(814307)
