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Abstract:

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.

Registro:

Documento: Artículo
Título:Rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by Escherichia coli and Clostridium sporogenes
Autor:Ertl, P.; Robello, E.; Battaglini, F.; Mikkelsen, S.R.
Filiación:Department of Chemistry, University of Waterloo, Waterloo, ON N2L 3G1, Canada
Departamento de Quimica Inorganica, Analitica y Quimica Fisica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Ciudad Universitaria, 1428 Buenos Aires, Argentina
Palabras clave: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
Año:2000
Volumen:72
Número:20
Página de inicio:4957
Página de fin:4964
DOI: http://dx.doi.org/10.1021/ac0003596
Título revista:Analytical Chemistry
Título revista abreviado:Anal. Chem.
ISSN:00032700
CODEN:ANCHA
CAS:2,6 dichlorophenolindophenol, 620-45-1, 956-48-9; bacitracin, 1405-87-4; cephalosporin C, 61-24-5; chloramphenicol, 134-90-7, 2787-09-9, 56-75-7; cycloserine, 339-72-0, 68-39-3, 68-41-7; erythromycin, 114-07-8, 70536-18-4; nalidixic acid, 389-08-2; nystatin, 1400-61-9, 34786-70-4, 62997-67-5; penicillin G, 1406-05-9, 61-33-6; platinum, 7440-06-4; potassium ferricyanide, 13746-66-2; rifampicin, 13292-46-1; silver, 7440-22-4; silver chloride, 7783-90-6; streptomycin, 57-92-1; succinic acid, 110-15-6; tetracycline, 23843-90-5, 60-54-8, 64-75-5, 8021-86-1; trimethoprim, 738-70-5; vancomycin, 1404-90-6, 1404-93-9; Anti-Bacterial Agents; Ferricyanides; hexacyanoferrate III, 13408-62-3
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00032700_v72_n20_p4957_Ertl

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Citas:

---------- APA ----------
Ertl, P., Robello, E., Battaglini, F. & Mikkelsen, S.R. (2000) . Rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by Escherichia coli and Clostridium sporogenes. Analytical Chemistry, 72(20), 4957-4964.
http://dx.doi.org/10.1021/ac0003596
---------- CHICAGO ----------
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" . Analytical Chemistry 72, no. 20 (2000) : 4957-4964.
http://dx.doi.org/10.1021/ac0003596
---------- MLA ----------
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" . Analytical Chemistry, vol. 72, no. 20, 2000, pp. 4957-4964.
http://dx.doi.org/10.1021/ac0003596
---------- VANCOUVER ----------
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. Anal. Chem. 2000;72(20):4957-4964.
http://dx.doi.org/10.1021/ac0003596