Rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by Escherichia coli and Clostridium sporogenes
Electrochemical measurement of respiratory chain activity allows rapid and reliable screening for antibiotic susceptibility in microorganisms. Chronoamperometry and chronocoulometry of suspensions of aerobically cultivated E. coli combined with the non-native oxidant potassium hexacyanoferrate(III)...
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todo:paper_00032700_v72_n20_p4957_Ertl2023-10-03T13:55:59Z Rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by Escherichia coli and Clostridium sporogenes Ertl, P. Robello, E. Battaglini, F. Mikkelsen, S.R. 2,6 dichlorophenolindophenol antibiotic agent bacitracin cephalosporin C chloramphenicol cycloserine erythromycin nalidixic acid nystatin penicillin G platinum potassium ferricyanide rifampicin silver silver chloride streptomycin succinic acid tetracycline trimethoprim vancomycin agar gel diffusion amperometry antibiotic sensitivity article bacterium culture Clostridium sporogenes coulometry electrochemistry Escherichia coli growth inhibition IC 50 measurement nonhuman reduction respiratory chain turbidimetry Anti-Bacterial Agents Clostridium Electrochemistry Escherichia coli Ferricyanides Microbial Sensitivity Tests Oxidation-Reduction Electrochemical measurement of respiratory chain activity allows rapid and reliable screening for antibiotic susceptibility in microorganisms. Chronoamperometry and chronocoulometry of suspensions of aerobically cultivated E. coli combined with the non-native oxidant potassium hexacyanoferrate(III) (ferricyanide) yield signals for reoxidation of the reduction product ferrocyanide that are much smaller if the E. coli has been incubated briefly with an effective antibiotic compound. Chronocoulometric results, obtained following 20-min incubation with antibiotic and 2-min measurement in assay buffer containing 50 mM ferricyanide and 10 mM succinate, at +0.50 V vs Ag/AgCl at a Pt working electrode, were compared with traditional disk diffusion susceptibility testing, which requires overnight incubation on agar plates; the results show significantly lower accumulation of ferrocyanide in all cases in which growth inhibition was observed in the disk diffusion assay. A range of antibiotic compounds (13) were examined that possess different mechanisms of action. Quantitative determination of IC50 values for penicilin G and chloramphenicol yielded values that were 100-fold higher than those obtained by standard turbidity methods after 10-h incubation; this is likely a result of the very brief (10 min) exposure time to the antibiotics. Addition of 5 μM 2,6-dichlorophenolindophenol, a hydrophobic electron-transfer mediator, to the assay mixture allowed susceptibility testing of a Gram-positive obligate anaerobe, Clostridium sporogenes. This rapid new assay will facilitate clinical susceptibility testing, allowing appropriate treatment virtually as soon as a clinical isolate can be obtained. Fil:Battaglini, F. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00032700_v72_n20_p4957_Ertl |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
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R-134 |
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
2,6 dichlorophenolindophenol antibiotic agent bacitracin cephalosporin C chloramphenicol cycloserine erythromycin nalidixic acid nystatin penicillin G platinum potassium ferricyanide rifampicin silver silver chloride streptomycin succinic acid tetracycline trimethoprim vancomycin agar gel diffusion amperometry antibiotic sensitivity article bacterium culture Clostridium sporogenes coulometry electrochemistry Escherichia coli growth inhibition IC 50 measurement nonhuman reduction respiratory chain turbidimetry Anti-Bacterial Agents Clostridium Electrochemistry Escherichia coli Ferricyanides Microbial Sensitivity Tests Oxidation-Reduction |
| spellingShingle |
2,6 dichlorophenolindophenol antibiotic agent bacitracin cephalosporin C chloramphenicol cycloserine erythromycin nalidixic acid nystatin penicillin G platinum potassium ferricyanide rifampicin silver silver chloride streptomycin succinic acid tetracycline trimethoprim vancomycin agar gel diffusion amperometry antibiotic sensitivity article bacterium culture Clostridium sporogenes coulometry electrochemistry Escherichia coli growth inhibition IC 50 measurement nonhuman reduction respiratory chain turbidimetry Anti-Bacterial Agents Clostridium Electrochemistry Escherichia coli Ferricyanides Microbial Sensitivity Tests Oxidation-Reduction Ertl, P. Robello, E. Battaglini, F. Mikkelsen, S.R. Rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by Escherichia coli and Clostridium sporogenes |
| topic_facet |
2,6 dichlorophenolindophenol antibiotic agent bacitracin cephalosporin C chloramphenicol cycloserine erythromycin nalidixic acid nystatin penicillin G platinum potassium ferricyanide rifampicin silver silver chloride streptomycin succinic acid tetracycline trimethoprim vancomycin agar gel diffusion amperometry antibiotic sensitivity article bacterium culture Clostridium sporogenes coulometry electrochemistry Escherichia coli growth inhibition IC 50 measurement nonhuman reduction respiratory chain turbidimetry Anti-Bacterial Agents Clostridium Electrochemistry Escherichia coli Ferricyanides Microbial Sensitivity Tests Oxidation-Reduction |
| description |
Electrochemical measurement of respiratory chain activity allows rapid and reliable screening for antibiotic susceptibility in microorganisms. Chronoamperometry and chronocoulometry of suspensions of aerobically cultivated E. coli combined with the non-native oxidant potassium hexacyanoferrate(III) (ferricyanide) yield signals for reoxidation of the reduction product ferrocyanide that are much smaller if the E. coli has been incubated briefly with an effective antibiotic compound. Chronocoulometric results, obtained following 20-min incubation with antibiotic and 2-min measurement in assay buffer containing 50 mM ferricyanide and 10 mM succinate, at +0.50 V vs Ag/AgCl at a Pt working electrode, were compared with traditional disk diffusion susceptibility testing, which requires overnight incubation on agar plates; the results show significantly lower accumulation of ferrocyanide in all cases in which growth inhibition was observed in the disk diffusion assay. A range of antibiotic compounds (13) were examined that possess different mechanisms of action. Quantitative determination of IC50 values for penicilin G and chloramphenicol yielded values that were 100-fold higher than those obtained by standard turbidity methods after 10-h incubation; this is likely a result of the very brief (10 min) exposure time to the antibiotics. Addition of 5 μM 2,6-dichlorophenolindophenol, a hydrophobic electron-transfer mediator, to the assay mixture allowed susceptibility testing of a Gram-positive obligate anaerobe, Clostridium sporogenes. This rapid new assay will facilitate clinical susceptibility testing, allowing appropriate treatment virtually as soon as a clinical isolate can be obtained. |
| format |
JOUR |
| author |
Ertl, P. Robello, E. Battaglini, F. Mikkelsen, S.R. |
| author_facet |
Ertl, P. Robello, E. Battaglini, F. Mikkelsen, S.R. |
| author_sort |
Ertl, P. |
| title |
Rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by Escherichia coli and Clostridium sporogenes |
| title_short |
Rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by Escherichia coli and Clostridium sporogenes |
| title_full |
Rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by Escherichia coli and Clostridium sporogenes |
| title_fullStr |
Rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by Escherichia coli and Clostridium sporogenes |
| title_full_unstemmed |
Rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by Escherichia coli and Clostridium sporogenes |
| title_sort |
rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by escherichia coli and clostridium sporogenes |
| url |
http://hdl.handle.net/20.500.12110/paper_00032700_v72_n20_p4957_Ertl |
| work_keys_str_mv |
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