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Journal of Clinical Microbiology, February 2006, p. 378-382, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.378-382.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Influence of Anti-FloR Antibody on Florfenicol Accumulation in Florfenicol-Resistant Escherichia coli and Enzyme-Linked Immunosorbent Assay for Detection of Florfenicol-Resistant E. coli Isolates

Beibei Wu,1 Chun Xia,2* Xiangdang Du,1 Xingyuan Cao,1 and Jianzhong Shen1*

Department of Pharmacology and Toxicology,1 Department of Microbiology and Immunology, College of Veterinary Medicine, China Agricultural University, Beijing 100094, People's Republic of China2

Received 2 May 2005/ Returned for modification 15 June 2005/ Accepted 16 November 2005

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.

* Corresponding author. Mailing address: Department of Pharmacology and Toxicology, College of Veterinary Medicine, China Agricultural University, Beijing 100094, People's Republic of China. Phone: 86-10-62732803. Fax: 86-10-62731032. E-mail: sjz{at}cau.edu.cn.

Journal of Clinical Microbiology, February 2006, p. 378-382, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.378-382.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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