上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (9): 1004-.doi: 10.3969/j.issn.1674-8115.2019.09.011

• 论著·基础研究 • 上一篇    下一篇

relA基因敲除对多黏菌素抗鲍曼不动杆菌异质性耐药的影响

余春波 1，卢明 2，邵雷 3，蒲甜 4，陈代杰 4，周薇 1, 5   

  1. 1. 上海交通大学医学院附属第九人民医院，上海 200011；2.中国医药工业研究总院 /上海医药工业研究院，上海 201203；3.上海健康医学院协同科研中心，上海 201318；4.上海交通大学药学院，上海 200240；5.上海交通大学医学院附属第九人民医院口腔医学院，口腔微生态与系统性疾病实验室，国家口腔疾病临床研究中心，上海市口腔医学重点实验室，上海市口腔医学研究所，上海 200011
  • 出版日期:2019-09-28 发布日期:2019-11-02
  • 通讯作者: 周薇，电子信箱：sweetzw@hotmail.com。
  • 作者简介:余春波（1995—），男，硕士生；电子信箱： ycb694722321@163.com。
  • 基金资助:
    上海市教育委员会高峰高原学科建设计划（20152523）

Effect of relA knockout on heterologous resistance of colistin to Acinetobacter baumannii

YU Chun-bo1, LU Ming2, SHAO Lei3, PU Tian4, CHEN Dai-jie4, ZHOU Wei1, 5   

  1. 1. Shanghai Ninth Peoples Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China; 2. China National Institute of Pharmaceutical Industry & Shanghai Institute of Pharmaceutical Industry, Shanghai 201203, China; 3. Shanghai Health Medical College, Collaborative research center, Shanghai 201318, China; 4. Shanghai Jiao Tong University School of Pharmacy, Shanghai 200240, China; 5. Shanghai Ninth Peoples Hospital, School of Stomatology, Shanghai Jiao Tong University School of Medicine; Laboratory of Oral Microbiota and Systemic Diseases; National Center for Clinical Research of Oral Diseases; Shanghai Key Laboratory of Stomatology & Shanghai Institute of Stomatology, Shanghai 200011, China
  • Online:2019-09-28 Published:2019-11-02
  • Supported by:
    Shanghai Municipal Education Commission—Gaofeng Clinical Medicine Support，20152523）。

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摘要： 目的 ·观察严谨反应相关的 (p)ppGpp合成酶基因（ relA）对多黏菌素抗鲍曼不动杆菌异质性耐药的影响。方法 ·采用 Red同源重组技术敲除鲍曼不动杆菌 ATCC19606中 relA基因；用结晶紫染色法观察鲍曼不动杆菌菌膜形成的情况；用群体分析法（population analysis profiles，PAP）检测在多黏菌素作用下鲍曼不动杆菌产生异质性耐药菌落数的变化并计算异质性耐药率；用杀菌曲线检测在多黏菌素作用下鲍曼不动杆菌持留菌形成情况。结果 ·成功敲除鲍曼不动杆菌中 relA基因，获得 relA基因敲除株 ATCC19606-ΔrelA，鲍曼不动杆菌的菌膜形成量明显下降，多黏菌素作用下异质性耐药菌落数及持留菌形成量均显著性降低。结论 ·细菌严谨反应中 (p)ppGpp合成酶 relA基因可能是影响鲍曼不动杆菌对多黏菌素产生异质性耐药的重要因素。

关键词: 鲍曼不动杆菌, relA基因, 多黏菌素, Red同源重组技术, 异质性耐药, 持留性, 菌膜

Abstract:

Objective · To observe the effect of the (p)ppGpp synthase gene (relA) on the heteroresistance of colistin against Acinetobacter baumannii. Methods · The relA gene in Acinetobacter baumannii ATCC19606 was knocked outRed homologous recombination technique. The biofilm formation of Acinetobacter baumannii was observedcrystal violet staining. The change of heterogeneous colonies of Acinetobacter baumannii under the action of colistin was detectedpopulation analysis profiles (PAP) and the heterogeneity was calculated. The killing curve was used to detect the formation of persistent bacteria in Acinetobacter baumannii under the action of colistin. Results · The relA gene in Acinetobacter baumannii was successfully knocked out, and the relA knockout strain ATCC19606-ΔrelA was obtained. After relA gene knockout, the biofilm formation of Acinetobacter baumannii decreased significantly. After relA gene knockout, Acinetobacter baumannii significantly reduced the heterogeneous colonies and persistent bacteria formation under the action of colistin. Conclusion · The bacterial stringent reaction (p) ppGpp synthase relA may be an important factor affecting the heteroresistance of Acinetobacter baumannii to colistin.

Key words: Acinetobacter baumannii, relA gene, colistin, Red homologous recombination technology, heteroresistance, persistence, biofilm

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