Please use this identifier to cite or link to this item: https://doi.org/10.1371/journal.pone.0140446
Title: Using an adenosine triphosphate bioluminescent assay to determine effective antibiotic combinations against carbapenem-resistant gram negative bacteria within 24 hours
Authors: Cai Y.
Leck H.
Lim T.P. 
Teo J.
Lee W.
Hsu L.Y. 
Koh T.H. 
Tan T.T. 
Tan T.-Y.
Kwa A.L.-H. 
Keywords: adenosine triphosphate
carbapenem
adenosine triphosphate
antiinfective agent
carbapenem derivative
Acinetobacter baumannii
antibiotic resistance
Article
bacterial count
bacterial strain
bioluminescence
colony forming unit
concentration (parameters)
controlled study
Gram negative bacterium
Klebsiella pneumoniae
measurement accuracy
nonhuman
prospective study
Pseudomonas aeruginosa
receiver operating characteristic
sensitivity and specificity
validation process
dose response
drug effects
Gram negative bacterium
luminescence
metabolism
microbial sensitivity test
multidrug resistance
procedures
reproducibility
Adenosine Triphosphate
Anti-Bacterial Agents
Carbapenems
Colony Count, Microbial
Dose-Response Relationship, Drug
Drug Resistance, Multiple, Bacterial
Gram-Negative Bacteria
Luminescent Measurements
Microbial Sensitivity Tests
Reproducibility of Results
ROC Curve
Issue Date: 2015
Citation: Cai Y., Leck H., Lim T.P., Teo J., Lee W., Hsu L.Y., Koh T.H., Tan T.T., Tan T.-Y., Kwa A.L.-H. (2015). Using an adenosine triphosphate bioluminescent assay to determine effective antibiotic combinations against carbapenem-resistant gram negative bacteria within 24 hours. PLoS ONE 10 (10) : e0140446. ScholarBank@NUS Repository. https://doi.org/10.1371/journal.pone.0140446
Abstract: Background Current in vitro combination testing methods involve enumeration by bacterial plating, which is labor-intensive and time-consuming. Measurement of bioluminescence, released when bacterial adenosine triphosphate binds to firefly luciferin-luciferase, has been proposed as a surrogate for bacterial counts. We developed an ATP bioluminescent combination testing assay with a rapid turnaround time of 24h to determine effective antibiotic combinations. Methods 100 strains of carbapenem-resistant (CR) GNB [30 Acinetobacter baumannii (AB), 30 Pseudomonas aeruginosa (PA) and 40 Klebsiella pneumoniae (KP)] were used. Bacterial suspensions (105 CFU/ml) were added to 96-well plates containing clinically achievable concentrations of multiple single and two-antibiotic combinations. At 24h, the luminescence intensity of each well was measured. Receiver operator characteristic curves were plotted to determine optimal luminescence threshold (TRLU) to discriminate between inhibitory/noninhibitory combinations when compared to viable plating. The unweighted accuracy (UA) [(sensitivity + specificity)/2] of TRLU values was determined. External validation was further done using 50 additional CR-GNB. Predictive accuracies of TRLU were high for when all antibiotic combinations and species were collectively analyzed (TRLU = 0.81, UA = 89%). When individual thresholds for each species were determined, UA remained high. Predictive accuracy was highest for KP (TRLU = 0.81, UA = 91%), and lowest for AB (TRLU = 0.83, UA = 87%). Upon external validation, high overall accuracy (91%) was observed. The assay distinguished inhibitory/noninhibitory combinations with UA of 80%, 94% and 93% for AB, PA and KP respectively. Conclusion We developed an assay that is robust at identifying useful combinations with a rapid turnaround time of 24h, and may be employed to guide the timely selection of effective antibiotic combinations. © 2015 Cai et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Source Title: PLoS ONE
URI: https://scholarbank.nus.edu.sg/handle/10635/161485
ISSN: 19326203
DOI: 10.1371/journal.pone.0140446
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