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Transference of methicillin resistance in Staphylococcus aureus using antibiotic challenge.

机译:使用抗生素攻击在金黄色葡萄球菌中耐甲氧西林的转移。

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摘要

Scope and method of study. The specific aim of this study was to document the transfer of a functional methicillin resistance gene, mecA , from methicillin resistant Staphylococcus aureus (MRSA) genomic DNA to methicillin sensitive Staphylococcal aureus (MSSA) thereby converting MSSA to MRSA. The experimental approach included the characterization of Staphylococcus aureus strains using Gram stain, coagulase and mannitol salt agar tests. Included among the Staphylococcus strains analyzed were 18 de-identified MRSA hospital isolates, nine MSSA strains isolated from band musical instruments and mouth-guards studies, and one MSSA strain of unknown origin. Additionally, sensitivity profiles of each strain were determined by minimum inhibitory concentration (MIC). Detection of mecA gene involved amplification by polymerase chain reaction (PCR) using fluorescently tagged primers and capillary electrophoresis with an ABI Prism Genetic Analyzer 310. Transformation studies involved the suspension of DNA extracted from two MRSA strains into a MSSA culture containing oxacillin complete with necessary controls. Electroporation was used in an attempt to force transformation, but it too was unsuccessful.;Findings and conclusions. Transformation was not observed under the specific test conditions. As a result of the electroporation experiment, growth was observed but no mecA gene was detected. The difficulty in transforming MSSA to MRSA suggests that transformation of SCCmec containing a mecA gene may not readily occur in nature or if so, it may proceed under conditions that were not replicated in vitro. Furthermore, additional analysis of SCCmec containing a mecA gene of the Staphylococcal strains may be necessary to further assess the integration of mecA into a recipient's genome. Lastly, transference of mecA to MSSA may be a result of conjugation or transduction, not transformation.
机译:研究范围和方法。这项研究的特定目的是证明功能性甲氧西林抗性基因mecA从耐甲氧西林的金黄色葡萄球菌(MRSA)基因组DNA转移至对甲氧西林敏感的金黄色葡萄球菌(MSSA),从而将MSSA转化为MRSA。实验方法包括使用革兰氏染色,凝固酶和甘露醇盐琼脂测试表征金黄色葡萄球菌菌株。在所分析的葡萄球菌菌株中,包括18种去鉴定的MRSA医院分离株,从乐队乐器和护口器研究中分离出的9种MSSA菌株以及一种来源不明的MSSA菌株。另外,通过最小抑制浓度(MIC)确定每个菌株的敏感性概况。 mecA基因的检测涉及通过荧光标记引物和ABI Prism Genetic Analyzer 310的毛细管电泳,通过聚合酶链反应(PCR)进行扩增。转化研究涉及将从两个MRSA菌株中提取的DNA悬浮到含有奥沙西林的MSSA培养物中,并进行必要的对照。电穿孔被用于试图强迫转化,但是也没有成功。;发现和结论。在特定的测试条件下未观察到转化。电穿孔实验的结果是观察到生长,但未检测到mecA基因。将MSSA转化为MRSA的困难表明,含有mecA基因的SCCmec转化在自然界可能不容易发生,或者如果这样,则可能在体外未复制的条件下进行。此外,可能需要对包含葡萄球菌菌株的mecA基因的SCCmec进行额外分析,以进一步评估mecA整合到受体基因组中的情况。最后,mecA向MSSA的转移可能是结合或转导而不是转化的结果。

著录项

  • 作者

    Pham, Trang Xuan.;

  • 作者单位

    Oklahoma State University.;

  • 授予单位 Oklahoma State University.;
  • 学科 Biology Microbiology.
  • 学位 M.S.
  • 年度 2011
  • 页码 78 p.
  • 总页数 78
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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