贵州医科大学学报

2016, v.41;No.186(03) 268-271

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11株临床诺卡菌16S rRNA及药敏分析
Identification and Drug Susceptibility Test of 11 Clinical Nocardia Strains

刘涛华;王颜颜;牟丽丽;吕倩;陈燕;张荣庆;康颖倩;
LIU Taohua;WANG Yanyan;MOU Lili;LYU Qian;CHEN Yan;ZHANG Rongqing;KANG Yingqian;Department of Microbiology,Guizhou Medical University;

摘要(Abstract):

目的:探讨应用11株临床诺卡菌16S rRNA序列鉴定该菌菌种名,同时观察该菌的药敏情况。方法:将11株临床分离的诺卡菌菌株分别接种于脑心浸液琼脂培养基分离培养,观察其生长情况;提取菌株DNA,聚合酶链式反应(PCR)扩增其16S rRNA序列,并测定其基因序列,所获序列与Gen Bank数据库进行BLAST比对,鉴别其菌种名;采用Kirby-Bauey纸片扩散法检测11株菌株药敏情况。结果:11株临床诺卡菌在脑心浸液琼脂培养基上生长迅速;PCR扩增菌株16S rRNA基因序列经BLAST比对,结果与Gen Bank中已知的诺卡.菌同源性>98%,分离的菌株分别为Nocardia otitidiscaviarum 4株、Nocardia cyriacigeorgica 3株、Nocrdia wallacei1株及Nocardia farcinica3株;药敏实验结果显示11株诺卡菌均对阿米卡星敏感,对亚胺培南耐药,其中Nocardia cyriacigeorgica对氨苄西林、环丙沙星、克拉霉素及阿莫西林的药物敏感性结果与其它菌株截然不同。结论:测定16S rRNA序列能够快速、简便、准确地鉴定诺卡菌菌种,各菌株均对阿米卡星敏感,对亚胺培南耐药。
Objective: To investigate the characteristics and susceptibility of the Nocardia by molecular genotyping method. Methods: Eleven clinical strains were inoculated in Brain Heart Infusion Agar medium separately and growth condition was observed. DNA was extracted and sequenced by PCR.Tested sequence was compared with Gen Bank Database by BLAST and identified its species name. The drug susceptibility of 11 strains was tested by Kirby- Bauev method. Results: The strains grew rapidly in Brain Heart Infusion Agar medium. The DNA homology of 16 S rRNA genes compared with the Nocardia strain known in Gen Bank were higher than 98% repectively. The collected clinical strains were Nocardia otitidiscaviarum( 4 strains),Nocardia cyriacigeorgica( 3 strains),Nocardia wallacei( 1strain) and Nocardia farcinica( 3 strains). All these strains were sensitive to amikacin and resistant to imipenem. Of which,Nocardia cyriacigeorgica demonstrated complete different drug susceptibility from other strains against ampicillin,ciprofloxacin,clarithromycin and amoxicillin. Conclusion: DNA sequencing analysis is a fast,simple,and accurate way to identify Nocardia. Tested strains are susceptible to amikacin and resistant to imipenem.

关键词(KeyWords): 诺卡菌;放线菌;16S rRNA;聚合酶链式反应;序列分析;DNA;药敏试验
Nocardia;actinomyces;16S rRNA;polymerase chain reaction;sequence analysis;DNA;drug susceptibility test

Abstract:

Keywords:

基金项目(Foundation): 国家自然基金地方项目(31060006,31260029);; 贵州省社会发展科技攻关项目[黔科合SY字(2011)3017号];; 贵州省卫生厅科技项目(gzwkj2010-1-025);; 贵阳市科技局社会发展与民生计划(筑科合同[2011103]16号)

作者(Author): 刘涛华;王颜颜;牟丽丽;吕倩;陈燕;张荣庆;康颖倩;
LIU Taohua;WANG Yanyan;MOU Lili;LYU Qian;CHEN Yan;ZHANG Rongqing;KANG Yingqian;Department of Microbiology,Guizhou Medical University;

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DOI: 10.19367/j.cnki.1000-2707.2016.03.007

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