Department of Clinical Laboratory, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, PR China.
Mol Med Rep. 2013 Mar;7(3):719-24. doi: 10.3892/mmr.2013.1284. Epub 2013 Jan 21.
The aim of this study was to detect class Ⅰ, II and III integrons using multiplex PCR, and to analyze the role that these integrons play in mediating multidrug-resistant Staphylococcus aureus (SA). The sensitivity of SA to 20 types of antibiotics was examined using the K-B method. A genomic DNA extraction kit was used for extracting genomic DNA and a high-purity 96 plasmid extraction kit was used for extracting plasmid DNA. Class Ⅰ, II and III integrons were amplified using multiplex PCR. Agarose gel electrophoresis was used for analysing amplification products. The positive rate of class Ⅰ and II integrons in the plasmid DNA from SA was higher compared to that of the genomic DNA. The positive rate of class Ⅰ integrons was highest in the group with multidrug resistance to amoxicillin/clavulanic acid, piperacillin/tazobactam, ciprofloxacin, tetracycline, rifampin, imipenem, cefazolin, cefuroxime, levofloxacin and gentamicin. As regards integron detection in the plasmids from drug-resistant SA strians obtained from sputum, blood, cerebrospinal fluid, drainage fluid, excretion and urine specimens, the difference in the detection rate of class Ⅰ integrons among the six types of specimens was significant. Multiplex PCR is an effective method to detect class Ⅰ, II and III integrons. The SA plasmid is the main carrier transferring integrons. Integrons mediate the formation of SA multidrug resistance.
本研究旨在通过多重 PCR 检测Ⅰ类、Ⅱ类和Ⅲ类整合子,并分析这些整合子在介导耐多药金黄色葡萄球菌(SA)中的作用。采用 K-B 法检测 SA 对 20 种抗生素的敏感性。使用基因组 DNA 提取试剂盒提取基因组 DNA,使用高纯度 96 质粒提取试剂盒提取质粒 DNA。采用多重 PCR 扩增Ⅰ类、Ⅱ类和Ⅲ类整合子。琼脂糖凝胶电泳分析扩增产物。与基因组 DNA 相比,SA 质粒中Ⅰ类和Ⅱ类整合子的阳性率更高。对阿莫西林/克拉维酸、哌拉西林/他唑巴坦、环丙沙星、四环素、利福平、亚胺培南、头孢唑林、头孢呋辛、左氧氟沙星和庆大霉素耐药的 SA 株的质粒中,Ⅰ类整合子的阳性率最高。在痰液、血液、脑脊液、引流液、排泄物和尿液标本中耐药 SA 株质粒中整合子的检测率,Ⅰ类整合子在 6 种标本中的检测率差异有统计学意义。多重 PCR 是检测Ⅰ类、Ⅱ类和Ⅲ类整合子的有效方法。SA 质粒是携带整合子的主要载体。整合子介导 SA 多药耐药的形成。