4.7 Article

Employing bacteria machinery for antibiotic detection: Using DNA gyrase for ciprofloxacin detection

Journal

CHEMICAL ENGINEERING JOURNAL
Volume 409, Issue -, Pages -

Publisher

ELSEVIER SCIENCE SA
DOI: 10.1016/j.cej.2020.128135

Keywords

Biochemical target of antibiotics; DNA gyrase; Ciprofloxacin; Screen-printed electrodes; Electrochemical biosensor; Antibodies

Funding

  1. European funds, through FEDER (European Funding or Regional Development) via COMPETE2020 - POCI (operational program for internationalization and competitively) [PTDC/AAG-TEC/5400/2014]
  2. National Foundation for Science and Technology, I.P. (FCT)
  3. National Foundation for Science and Technology, I.P. [SFRH/BD/130107/2017]
  4. Fundação para a Ciência e a Tecnologia [PTDC/AAG-TEC/5400/2014, SFRH/BD/130107/2017] Funding Source: FCT

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This study describes a successful new approach for designing biosensors that can detect antibiotics. It utilizes a biomimetic strategy by using the biochemical target of a given antibiotic as its biorecognition element. The biosensor showed sensitive responses for quinolones, particularly ciprofloxacin and norfloxacin, with a wide response range and selective response against the antibiotic group.
This work describes a new successful approach for designing biosensors that detect antibiotics. It makes use of a biomimetic strategy, by employing the biochemical target of a given antibiotic as its biorecognition element. This principle was tested herein for quinolones, which target DNA gyrase in bacteria. Ciprofloxacin (CIPRO) was tested as a representative antibiotic from the quinolone group; the sensitivity of biosensor to this group was confirmed by checking the response to another quinolone antibiotic (norfloxacin, NOR) and to a non-quinolone antibiotic (ampicillin, AMP). The biorecognition element used was DNA gyrase attached by ionic interactions to a carbon support, on a working electrode on common screen-printed electrodes (SPEs). The response against antibiotics was tested for increasing concentrations of CIPRO, NOR or AMP, and following the subsequent electrical changes by electrochemical impedance spectroscopy. The DNAgyrase biosensor showed sensitive responses for CIPRO and NOR, for concentrations down to 3.02 nM and 30.2 nM, respectively, with a very wide response range for CRIPRO, up to 30.2 mu M. Its response was also confirmed selective for quinolones, when compared to its response against AMP. Further comparison to an immunosensor of similar design (adding antibodies instead of DNA gyrase) was made, revealing favourable features for the new biomimetic biosensor with 1.52 nM of limit of detection (LOD). Overall, the new approach presented herein is simple and effective for antibiotic detection, displaying a selective response against a given antibiotic group. The use of bacterial machinery as biorecognition element in biosensors may also provide a valuable tool to study the mechanism of action in bacterial cells of new drugs. This is especially important in the development of new drugs to fight bacterial resistance.

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