首都医科大学学报 ›› 2013, Vol. 34 ›› Issue (5): 727-732.doi: 10.3969/j.issn.1006-7795.2013.05.019

• 临床研究 • 上一篇    下一篇

医院内宽嗜性铜绿假单胞菌噬菌体的分离筛选

代芳芳1, 赵秀英2, 于艳华1, 张立丽1, 陈铭1, 袁星星1, 李宁1   

  1. 1. 首都医科大学附属北京佑安医院临床检验中心, 北京 100069;
    2. 清华大学北京清华医院检验科, 北京 102218
  • 收稿日期:2013-06-14 出版日期:2013-10-21 发布日期:2013-10-22
  • 通讯作者: 赵秀英 E-mail:zhaoxiuying2001@163.com
  • 基金资助:

    北京市科技新星项目(2007B074);"215"高层次卫生技术人才队伍建设工程(2009-3-19);北京天坛医院青年基金(KY2011-03)。

Isolation and identification of pseudomonas aeruginosa phages with broad host range in hospital

DAI Fangfang1, ZHAO Xiuying2, YU Yanhua1, ZHANG Lili1, CHEN Ming1, YUAN Xingxing1, LI Ning1   

  1. 1. Department of Clinical Laboratory, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China;
    2. Department of Clinical Laboratory, Tsinghua Hospital, Tsinghua University, Beijing 102218, China
  • Received:2013-06-14 Online:2013-10-21 Published:2013-10-22
  • Supported by:

    This study was supported by Beijing Nova Program (2007B074), Beijing Municipal Health System "215" High-level Medical Personnel Culturing Project (2009-3-19), Youth Foundation of Beijing Tiantan Hospital (KY2011-03).

摘要:

目的 从未经处理的医院下水道污水中分离筛选宽嗜性铜绿假单胞菌噬菌体,检测其对临床常规检出的铜绿假单胞菌的嗜性覆盖范围,探究其潜在的临床治疗价值。方法 用双层琼脂噬菌斑形成法对铜绿假单胞菌噬菌体进行分离鉴定和感染谱的筛选,并用重复片段PCR基因指纹方法 (repetitive element PCR genomic fingerprinting, Rep-PCR)进行感染谱内宿主菌同源性分析,确认噬菌体的感染谱;用双层琼脂噬菌斑形成法检测筛出的宽嗜性铜绿假单胞菌噬菌体对临床常规检出的铜绿假单胞菌的嗜性覆盖范围。结果 从16家医院未经处理的下水道污水中分离到14株铜绿假单胞菌噬菌体,确定其中4株感染谱较广,4株宽嗜性噬菌体的交叉组合对临床日常分离30株铜绿假单胞菌吞噬覆盖率为96.7%(29/30);30株铜绿假单胞菌中对亚胺培南耐药菌株12株,对药敏实验全部药物敏感菌株5株,其他耐药菌株13株,4株宽嗜性噬菌体交叉组合对耐亚胺培南菌株吞噬覆盖率为91.7%(11/12),对药敏实验敏感菌株的吞噬覆盖率为100%(5/5),对其他耐药菌株交叉吞噬覆盖率为100%(13/13)。结论 从分离鉴定的14株铜绿假单胞菌噬菌体中筛选出4株宽嗜性噬菌体,其交叉组合能够吞噬型解临床日常筛出30株铜绿假单胞菌中的29株,对耐亚胺培南和其他临床常规药物的耐药菌株的交叉吞噬覆盖率在90%以上,交叉组合吞噬覆盖率高,为其进一步在临床抗菌治疗方面的应用提供基础和依据。

关键词: 宽嗜性, 铜绿假单胞菌噬菌体, 铜绿假单胞菌, 耐药菌株

Abstract:

Objective To isolate, screen pseudomonas aeruginosa phages with broad host range from hospital sewage wastewater and test their host spectrum to daily clinical isolates of pseudomonas aeruginosa to explore the application value in clinical antibiotic therapy. Methods pseudomonas aeruginosa phages were isolated and identified with plague method and their host spectrum were screened using the same method, the homology of the host bacteria were analysed by Rep-PCR in order to confirm the host spectrum at gene level; The lysis coverage of the combination of the broad host range phages were tested using plague method. Results Fourteen pseudomonas aeruginosa phages were isolated from sewage wastewater of sixteen hospitals, among the fourteen phages there were four phages with broad host range. The lysis coverage rate of the combination of the four broad host range phages to thirty pseudomonas aeruginosa strains isolated from daily test was 96.7%, to the twelve Imipenem-resistant ones in the thirty pseudomonas aeruginosa strains was 91.7%, to all the drug susceptive ones was 100%, and to other drug resistant ones was 100%. Conclusion Four pseudomonas aeruginosa phages with broad host range were isolated from fourteen pseudomonas aeruginosa phages. The combination of the four broad host range phages can lyse twenty-nine in thirty strains isolated from daily clinical test, the lysis coverage rate of the combination of the four broad host range phages to Imipenem-resistant pseudomonas aeruginosa and other drug resistant pseudomonas aeruginosa was above 90%, the combination had a high lysis coverage rate and this can lay the foundation and provide feasibility for clinical antibiotic therapy.

Key words: broad host range, pseudomonas aeruginosa phages, pseudomonas aeruginosa, drug resistant strains

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