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5株万古霉素敏感性减低耐甲氧西林金黄色葡萄球菌的graS基因及细胞超微结构分析

  

  1. 1.秦皇岛市海港医院 检验科,河北 秦皇岛 066000;2.河北医科大学第二医院 检验科  河北省临床检验中心,河北 石家庄 050000;
    3.华南理工大学轻工与食品学院,广东 广州  51064
  • 出版日期:2017-05-05 发布日期:2017-05-05
  • 通讯作者: 通信作者:赵建宏,Email: zhaojh_2002@yahoo.com
  • 基金资助:
    国家科技基础条件平台工作重点项目(2005DKA212026);河北省卫生厅医学重点课题(08075);河北医科大学第二医院重点课题(2h0200805)

graS gene and cell ultrastructure analysis of five methicillinresistant  Staphylococcus aureus with reduced susceptibility to vancomycin

  1. 1.Department of Laboratory Medicine,Qinhuangdao Haigang Hospital, Qinhuangdao 066000, China;
    2.Department of Laboratory Medicine, the Second Hospital  of  Hebei Medical University,
    Hebei Provincial Center for Clinical Laboratory, Shijiazhuang 050000, China;
    3.College of Light   Industry and  Food Sciences,South China University of  Technology, Guangzhou 510640,China
  • Online:2017-05-05 Published:2017-05-05
  • Contact: Corresponding author: Zhao Jianhong,Email: zhaojh_2002@yahoo.com

摘要: 目的分析在防耐药变异浓度(mutant prevention concentration, MPC)测定过程中筛选出的对万古霉素敏感性减低的耐甲氧西林金黄色葡萄球菌(methicillinresistant  Staphylococcus aureus, MRSA)在graS基因及细胞超微结构上的变化。 方法利用SauPCR、脉冲场凝胶电泳(pulsed field gel electrophoresis,PFGE)对筛选菌株进行分析。扩增graS基因,并利用变性梯度凝胶电泳 (denaturing gradient gel electrophoresis, DGGE)技术检测在该基因上是否存在突变。通过电子显微镜技术观察细胞壁结构改变。结果万古霉素敏感性减低菌株经SauPCR和PFGE分型检测,证实部分筛选菌株电泳条带与原始菌株存在有差异,且电镜结果也显示筛选菌株细胞壁有所增厚。DGGE检测未发现在graS基因上存在突变点。结论试验中的菌株万古霉素敏感性的减低可能不是由graS基因突变造成的,可能存在其他基因位点的突变,或可能存在另外的耐药机制。

关键词: 抗菌药;葡萄球菌, 金黄色;耐药性, 细菌

Abstract: ObjectiveThe methicillinresistant  Staphylococcus aureus (MRSA) with reduced susceptibility  to vancomycin which were selected  during  mutant prevention concentration(MPC) determination were characterized to compare the changes  in  the  graS  gene  and  cell  ultrastructure.MethodsThe strains selected  were characterized by pulsedfield gel  electrophoresis  (PFGE) and SauPCR  assay. The graS gene was detected by polymerase chain reaction method; Denaturing gradient gel electrophoresis (DGGE) was used to probe for mutations in the graS gene. The electron  microscopy was used to compare the differences on cell   wall.ResultsCompared with the parental strains, some strains with reduced susceptibility to vancomycin had different DNA profiles by SauPCR assay and PFGE. Electron  microscopy showed  that  the mutant  strains had  slightly  thicker cell walls  than  those of the wildtype  strains. DDGE was used to probe for mutations  in  the graS gene, but no different banding patterns were observed on the electrophoretogram  spectrum. ConclusionThe strains with reduced susceptibility to vancomycin were not caused by genetic mutations of graS, there may be mutations in genes other than the  graS gene or existence of other resistant mechanisms.

Key words: antibacterial agents, Staphylococcus aureus;drug resistance, bacterial