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Journal of Clinical Microbiology, April 2008, p. 1213-1219, Vol. 46, No. 4
0095-1137/08/$08.00+0     doi:10.1128/JCM.02036-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Rapid Antimicrobial Susceptibility Determination of Uropathogens in Clinical Urine Specimens by Use of ATP Bioluminescence

Vesna Ivani,¹ Mitra Mastali,² Neil Percy,³ Jeffrey Gornbein,⁴ Jane T. Babbitt,² Yang Li,¹ Elliot M. Landaw,⁴ David A. Bruckner,⁵ Bernard M. Churchill,¹ and David A. Haake^2,6*

Departments of Urology,¹ Medicine,² Biomathematics,⁴ Pathology and Laboratory Medicine, David Geffen School of Medicine at the University of California-Los Angeles, Los Angeles, California 90095,⁵ Hygiena, Camarillo, California 93012,³ Veterans Affairs Greater Los Angeles Healthcare System, Los Angeles, California 90073⁶

Received 17 October 2007/ Returned for modification 26 November 2007/ Accepted 4 February 2008

We describe the first direct testing of the antimicrobial susceptibilities of bacterial pathogens in human clinical fluid samples by the use of ATP bioluminescence. We developed an ATP bioluminescence assay that eliminates somatic sources of ATP to selectively quantify the bacterial load in clinical urine specimens with a sensitivity of <1,000 CFU per milliliter. There was a log-log relationship between light emission and the numbers of CFU in clinical urine specimens. A clinical study was performed to evaluate the accuracy of the ATP bioluminescence assay for determination of the antimicrobial susceptibilities of uropathogens in clinical urine specimens tested in a blinded manner. ATP bioluminescent bacterial density quantitation was used to determine the inoculation volume in growth medium with and without antibiotics. After incubation at 37°C for 120 min, the ATP bioluminescence assay was repeated to evaluate the uropathogen response to antibiotics. The ability of the ATP bioluminescence assay to discriminate between antimicrobial susceptibility and resistance was determined by comparison of the results obtained by the ATP bioluminescence assay with the results obtained by standard clinical microbiology methods. Receiver operator characteristic curves were used to determine the optimal threshold for discriminating between susceptibility and resistance. Susceptibility and resistance were correctly predicted in 87% and 95% of cases, respectively, for an overall unweighted accuracy of 91%, when the results were stratified by antibiotic. For samples in which the pathogen was susceptible, the accuracy improved to 95% when the results for samples with less than a 25-fold increase in the amount of bacterial ATP in the medium without antibiotics were excluded. These data indicate that a rapid bioluminescent antimicrobial susceptibility assay may be useful for the management of urinary tract infections.

Published ahead of print on 13 February 2008.