Wu Beibei, Xia Chun, Du Xiangdang, Cao Xingyuan, Shen Jianzhong
Department of Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University, Beijing 100094, People's Republic of China.
J Clin Microbiol. 2006 Feb;44(2):378-82. doi: 10.1128/JCM.44.2.378-382.2006.
To detect florfenicol-resistant Escherichia coli isolates by enzyme-linked immunosorbent assay (ELISA), anti-FloR1 antibodies were produced in mice using a recombinant glutathione S-transferase (GST)-FloR1 protein, which was expressed in a prokaryote expression system, as the antigen. The specificity of the murine anti-GST-FloR1 antibody and its influence on florfenicol accumulation in florfenicol-resistant isolates were investigated using Western blotting and high-performance liquid chromatography, respectively. Western blotting using the anti-FloR1 antibody showed specific binding of the antibody to the florfenicol-resistant FloR protein. Preincubation of florfenicol-resistant strains with the antibody significantly increased the intracellular accumulation of florfenicol and enhanced the bacterial susceptibility to florfenicol, suggesting that antibody binding to the FloR protein inhibited the activity of the efflux protein conferred by the floR gene. Analyses of florfenicol-resistant and -sensitive isolates by ELISA using the anti-FloR1 antibody showed good correlation between FloR protein expression and the floR genotype. The anti-FloR1 antibody-based ELISA is a useful tool for the detection of florfenicol-resistant bacteria harboring the floR gene.
为了通过酶联免疫吸附测定（ELISA）检测氟苯尼考耐药性大肠杆菌分离株，使用在原核表达系统中表达的重组谷胱甘肽S-转移酶（GST）-FloR1蛋白作为抗原，在小鼠体内产生抗FloR1抗体。分别使用蛋白质印迹法和高效液相色谱法研究了鼠抗GST-FloR1抗体的特异性及其对氟苯尼考耐药分离株中氟苯尼考积累的影响。使用抗FloR1抗体进行的蛋白质印迹显示该抗体与氟苯尼考耐药的FloR蛋白特异性结合。用该抗体对氟苯尼考耐药菌株进行预孵育可显著增加氟苯尼考在细胞内的积累，并增强细菌对氟苯尼考的敏感性，这表明抗体与FloR蛋白的结合抑制了floR基因赋予的外排蛋白的活性。使用抗FloR1抗体通过ELISA对氟苯尼考耐药和敏感分离株进行分析，结果显示FloR蛋白表达与floR基因型之间具有良好的相关性。基于抗FloR1抗体的ELISA是检测携带floR基因的氟苯尼考耐药细菌的有用工具。
