Rapid optical determination of β-lactamase and antibiotic activity

BMC Microbiology

Tayyaba Hasan²  Gerard J Nau¹  Xiang Zheng⁴  Ulysses W Sallum³  Shazia Khan² 

[1] Department of Microbiology and Molecular Genetics, Department of Medicine, Division of Infectious Diseases, Centre for Vaccine Research, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA;Wellman Centre for Photomedicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, USA;Present address: Partners Research Ventures and Licensing, 101 Huntington Ave., 4th Floor, Boston, MA 02199, USA;Present address: Pfizer Inc, 401 N Middletown Rd, Pearl River, NY 10965, USA

关键词: Antibiotic susceptibility;    Antibiotic activity;    β-lactam;    β-lactamase;    Cefazolin;    Cephalosporin;    Staphylococcus;    Quenching;    Fluorophore;    Fluorescence;   

Others  :  1141449
DOI  :  10.1186/1471-2180-14-84

 received in 2013-12-13, accepted in 2014-03-28,  发布年份 2014

【 摘 要 】

Background

The absence of rapid tests evaluating antibiotic susceptibility results in the empirical prescription of antibiotics. This can lead to treatment failures due to escalating antibiotic resistance, and also furthers the emergence of drug-resistant bacteria. This study reports a rapid optical method to detect β-lactamase and thereby assess activity of β-lactam antibiotics, which could provide an approach for targeted prescription of antibiotics. The methodology is centred on a fluorescence quenching based probe (β-LEAF – β-Lactamase Enzyme Activated Fluorophore) that mimics the structure of β-lactam antibiotics.

Results

The β-LEAF assay was performed for rapid determination of β-lactamase production and activity of β-lactam antibiotic (cefazolin) on a panel of Staphylococcus aureus ATCC strains and clinical isolates. Four of the clinical isolates were determined to be lactamase producers, with the capacity to inactivate cefazolin, out of the twenty-five isolates tested. These results were compared against gold standard methods, nitrocefin disk test for β-lactamase detection and disk diffusion for antibiotic susceptibility, showing results to be largely consistent. Furthermore, in the sub-set of β-lactamase producers, it was demonstrated and validated that multiple antibiotics (cefazolin, cefoxitin, cefepime) could be assessed simultaneously to predict the antibiotic that would be most active for a given bacterial isolate.

Conclusions

The study establishes the rapid β-LEAF assay for β-lactamase detection and prediction of antibiotic activity using S. aureus clinical isolates. Although the focus in the current study is β-lactamase-based resistance, the overall approach represents a broad diagnostic platform. In the long-term, these studies form the basis for the development of assays utilizing a broader variety of targets, pathogens and drugs.

【 授权许可】

   
2014 Khan et al.; licensee BioMed Central Ltd.

【 图 表 】

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