天津医药 ›› 2018, Vol. 46 ›› Issue (3): 246-250.doi: 10.11958/20171492

• 实验研究 • 上一篇    下一篇

TEM-1型β-内酰胺酶及CarO蛋白介导的耐舒巴坦鲍曼不动杆菌临床株耐药机制研究

李楠,张豪杰,王悦,祁萌,郭文学,王哲,祁伟△   

  1. 天津医科大学第二医院感染性疾病研究所(邮编300211)
  • 收稿日期:2017-12-22 修回日期:2018-01-22 出版日期:2018-03-15 发布日期:2018-03-23
  • 通讯作者: 李楠 E-mail:1450723549@qq.com

Study on the drug resistance of clinical strain of acinetobacter baumannii mediated by TEM-1 beta-lactamases and porin CarO

LI Nan, ZHANG Hao-jie, WANG Yue, Qi Meng, GUO Wen-xue, WANG Zhe, QI Wei△   

  1. Infectious Disease Institute, the Second Hospital of Tianjin Medical University, Tianjin 300211, China
  • Received:2017-12-22 Revised:2018-01-22 Published:2018-03-15 Online:2018-03-23
  • Contact: Nan LI E-mail:1450723549@qq.com

摘要: 目的 研究 TEM-1 型 β-内酰胺酶及 CarO 蛋白介导的鲍曼不动杆菌临床株对舒巴坦的耐药机制。方法 将 24 株非重复鲍曼不动杆菌临床株通过琼脂稀释法分为舒巴坦非敏感组(18 株)和敏感组(6 株),用纸片扩散法(K-B)行药敏试验,PCR 扩增 blaTEM-1和 carO 基因,选取 4 株 blaTEM-1阳性菌株(A3、A5、1327、C1),对其扩增产物测序,BLAST 软件分析;应用实时荧光定量 RT-PCR 技术检测 2 组菌株 blaTEM-1和 carO 基因 mRNA 转录情况。结果敏感组临床株对美罗培南、亚胺培南、头孢哌酮、环丙沙星、庆大霉素、氨苄西林耐药率分别为 1/6、2/6、3/6、1/6、5/6、5/6;非敏感组分别为 6/18、10/18、18/18、18/18、18/18。敏感组未检出 blaTEM-1,非敏感组中 16 株临床株扩增出 blaTEM-1;24 株临床菌株全部扩增出 carO 基因。测序显示 4 株临床株 blaTEM-1基因均未出现有义突变;A3、A5、1327 启动序列为 P4,C1 为 P3。BlaTEM-1基因阳性菌株 mRNA 相对表达量与其对舒巴坦 MIC 值呈正相关(rs=0.551,P=0.027);carO 基因 mRNA 相对表达在敏感组和非敏感组中无差异。结论 临床分离鲍曼不动杆菌受试菌株耐药严重,其对舒巴坦的耐药机制与 TEM-1 型 β-内酰胺酶高表达有关,而 blaTEM-1基因启动子调控可能是 TEM-1 型 β-内酰胺酶高表达的原因之一。

关键词: 鲍氏不动杆菌, β 内酰胺酶类, 微生物敏感性试验, carO 基因, blaTEM-1基因, TEM-1 型 β-内酰胺酶

Abstract: Objective To investigate the mechanism of drug resistance of sulbactam mediated by TEM-1 betalactamases and porin CarO in clinical strains of acinetobacter baumannii. Methods Twenty-four unrepeated clinical acinetobacter baumannii strains were divided into sensitive strain group (n=6) and insensitive strain group (n=18) by susceptibility testing to sulbactam. Antibiotics susceptibility test was carried out using the Kirby-Bauer method. BlaTEM-1 and carO genes were amplified by PCR. Four blaTEM-1 positive strains (A3, A5, 1327 and C1) were selected, and their amplified products were sequenced. The quantitative real-time RT-PCR was used to analyze mRNA transcriptional levels of blaTEM-1 and carO genes. Results Resistant rates of the sensitive strain group for meropenem, imipenem, cefoperazone,ciprofloxacin, gentamicin and ampicillin were 1/6, 2/6, 3/6, 1/6, 5/6 and 5/6, and resistant rates of the insensitive strain group were 6/18, 10/18, 18/18, 18/18 and 18/18. BlaTEM-1 genes were amplified in 16 insensitive strains, and blaTEM-1 was negative in sensitive strains. The carO genes were amplified in all 24 strains. There was no significative mutation in the 4 strains of blaTEM-1 genes. The promoters of the strains A3, A5 and 1327 were P4, and C1 was P3. There was a positive correlation between the mRNA expression of blaTEM-1 and the MIC value of sulbactam (rs=0.551, P=0.027). There was no difference in the mRNA expression of carO between the two groups. Conclusion The clinical strains are seriously resistant to antibiotics. The main resistance mechanism of clinical strains to sulbactam is the high mRNA expression of blaTEM-1, and the promoter may be one of the reasons of high expression of TEM-1

Key words: Acinetobacter baumannii, beta-Lactamases, microbial sensitivity tests, carO gene, blaTEM-1 gene, TEM-1 β-lactamase