革兰阴性杆菌碳青霉烯酶产生与细菌耐药性关系的研究  

The relationship between acquired carbapenemases and resistance of gram-negative bacill

作　　者：沈继录[1] 朱德妹[1] 吴卫红[1] 徐晓刚[1] 王明贵[1] 

机构地区：[1]复旦大学附属华山医院抗生素研究所,上海200040

出　　处：《中华检验医学杂志》2008年第4期408-414,共7页Chinese Journal of Laboratory Medicine

基　　金：基金项目：国家重点基础研究发展计划（973计划）资助项目（2005CB523101）

摘　　要：目的 研究耐亚胺培南革兰阴性杆菌碳青霉烯酶产生及流行情况。方法采用琼脂稀释法测定亚胺培南和美罗培南对199株革兰阴性杆菌的MIC，采用乙二胺四乙酸（EDTA）协同实验筛选金属β内酰胺酶表型。PCR扩增耐药菌株碳青霉烯酶基因，并测序分析。脉冲场凝胶电泳（PFGE）分析产酶菌株同源性。结果 141株耐碳青霉烯类铜绿假单胞菌对亚胺培南和美罗培南药敏试验结果显示具有3种模式，以亚胺培南和美罗培南同时耐药为主94株（66．7％）、亚胺培南耐药和美罗培南敏感46株（32．6％）、亚胺培南敏感和美罗培南耐药仅1株（0．7％）。但其他耐碳青霉烯类的鲍曼不动杆菌、洛菲不动杆菌、弗劳地柠檬酸杆菌、肺炎克雷伯菌和黏质沙雷菌均对亚胺培南和美罗培南同时耐药。EDTA协同试验结果显示，仅4株耐亚胺培南铜绿假单胞菌为EDTA协同试验阳性（2．8％），其余菌株均为EDTA协同试验阴性。PCR扩增碳青霉烯酶基因结果显示，4株EDTA协同试验阳性的铜绿假单胞菌产VIM-2型金属酶；34株鲍曼不动杆菌菌株中30株（88．2％）产OXA型碳青霉烯酶，其中OXA23型27株（79．4％），OXA24型13株（38．2％），OXA66型23株（67．6％）。并且22株（64．7％）细菌同时产生一种以上的OXA型碳青霉烯酶。7株洛菲不动杆菌全部产OXA-23型碳青霉烯酶；11株弗劳地柠檬酸杆菌、5株肺炎克雷伯菌和1株黏质沙雷菌均产KPC-2型碳青霉烯酶，其中6株弗劳地柠檬酸杆菌同时具有IMP-8新亚型金属酶。PFGE结果显示，34株鲍曼不动杆菌中PFGE谱型共有15种，其中有14株属于A型，7株属于B型；7株洛菲不动杆菌不属于同一克隆；4株产VIM-2型金属酶铜绿假单胞菌不属于同一PFGE谱型；11株柠檬酸杆菌属于同一PFGE谱型；5株肺炎克雷伯菌属于同一PFGE谱型。结论耐碳青霉烯类革兰阴性杆菌对12种抗生素的耐药�Objective To investigate acquired carbapenemases and prevalence of carbapenemresistant gram-negative bacill. Methods The antimicrobial susceptibility was determined by agar dilution method. Metallo-β-lactamase（MBLs） were screened by EDTA-disk synergy test. The encoding genes of MBLs were amplified by PCR followed by sequencing. Strain homology was investigated by pulsed-field gel electronphoresis（PFGE）. Results In 141 carbapenem-resistant Pseudomonas aeruginosa（P. aeruginosa） , there were three resistant patterns which were imipenem （IMP） -resistant ＋ merooenem （ MEM ） -resistant （66. 7% ）, IMP-resistant ＋ MEM-sensitive （32. 6% ）, and IMP-resistant ＋ MEM-sensitive （0. 7% ）. All the carbapenem-resistan Acinetobacter baumannii （ A. baumannii）, Acinetobacter lwoffi （ A. lwoffi）, Citrobactor freundii（ C. freundii）, Klebsiella pneumoniae（ K. pneumoniae） and Serratia were resistant to imipenem and meropenem. Four strains of 141 P. aeruginosa were positive by EDTA-disk synergy test, and they produced VIM-2-type Metallo-β-1actamase. Of 34 carbapenem-resistan A. baumannii, 30 strains produced OXA- type. Among them, OXA23, OXA24 and OXA66 accounted for 79.4%, 38. 2% and 67. 6%, respectively. And 22 of 34 strains （64. 7% ）produced multiple OXA-carbapenemases. All 7 strains A. lwoffi produced OXA-23-type carbapenemases. All 11 strains C. freundii, 5 strains K. pneumoniae and 1 strains Serratia produced KPC-2-type carbapenemases. And 6 of 11 strains C. freundii produced new subtype IMP- S. Of 15 PFGE type in 34 strains A. baumannii, 14 strains belonged to A-type,7 strains belonged to Btype. Seven A. lwoffi strains distributed in different PFGE type. Four strains of P. aeruginosa producing VIM- 2-type Metallo-β-1actamase did not have the same PFGE type. Eleven C. freundii strains had the same PFGE type. Five K. pneumoniae strains had the same PFGE type. Conchmions Drug resistance to 12 common antibiotics in carbapenem-resistant gram-negative bacill was higher than no

关 键 词：革兰阴性菌 细胞蛋白质类 Β内酰胺酶类 抗药性 细菌 

分 类 号：R686[医药卫生—骨科学]