Rapid antibiotic susceptibility tests on Enterobacteriaceae by ATP bioluminescence

Wheat, P. F.; Hastings, James; Spencer, R C

doi:10.1099/00222615-25-2-95

Cited by 27 publications

(19 citation statements)

References 14 publications

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“…Bioluminescence methods have been shown to be useful for the rapid (2-to 4-h) determination of the antibiotic (15,27,31,32). The cutoff for susceptibility was determined to be Ͻ40% of the RLU measurement obtained for bacteria in the absence of antibiotic (15).…”

Section: Discussionmentioning

confidence: 99%

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

Ivančić¹,

Mastali²,

Percy³

et al. 2008

J Clin Microbiol

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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.The urinary tract is the most common organ system to experience bacterial infections (17). As a major cause of patient morbidity and health care expenditures for men and women of all age groups, urinary tract infections (UTIs) account for 7 million office visits, more than 1 million emergency room visits, and 100,000 hospitalizations each year (20). The estimated annual cost to the U.S. health care system is approximately $1.6 billion (8). Patients with nosocomial and/or complicated UTIs due to abnormal urinary tract anatomy, the presence of indwelling foreign bodies, and obstructed kidney stones are at risk for life-threatening systemic infections and permanently reduced kidney function. Patients with recurrent communityacquired, simple UTIs may incur significant treatment costs and substantial morbidity due to bothersome symptoms, such as painful urination, frequency, and urgency.The traditional basis for the evaluation of urinary tract pathogens (uropathogens) is urine culture and antibiotic susceptibility testing (13). The major drawback of the current microbiology approach is the time lapse of 2 to 3 days between s...

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Section: Discussionmentioning

confidence: 99%

“…The bacterial ATP bioluminescence methodology has been shown to be useful for the rapid (2-to 4-h) determination of the antibiotic susceptibilities of cultured clinical isolates (15,27,31,32). In this study, we evaluated the analytic validity of bioluminescent antibiotic susceptibility testing of uropathogens in clinical urine specimens.…”

mentioning

confidence: 99%

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

Ivančić¹,

Mastali²,

Percy³

et al. 2008

J Clin Microbiol

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“…However, in view of the similarities between bacterial and mammalian cytoplasmic membranes, the use of DiOC5(3) to monitor membrane potential changes in bacteria seems reasonable. Published applications of flow cytometry for studies of microorganisms using different parameter combinations cover a variety of fields that include microorganism identification, cell cycle analysis, and assessment of antibiotic action (2,9,12,14,15,16). Membrane potential measurements on bacteria have also been proposed as indicators of microbial viability and for bacterial detection, identification, and antibiotic susceptibility testing (13).…”

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confidence: 99%

Rapid flow cytometric antibiotic susceptibility assay for Staphylococcus aureus

Ordóñez

Wehman

1993

Cytometry

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Conventional techniques for antibiotic susceptibility testing usually require 24 h or more to produce accurate results. This long waiting period to demonstrate antibiotic action is necessary because such tests rely on growth (or the lack of it), when the microorganisms are incubated in the presence of the antibiotic. In an effort to improve antibiotic susceptibility testing, we developed a flow cytometric technique for Staphylococcus aureus in which antibiotic action is determined by monitoring druginduced membrane potential changes, using the potential-sensitive dye 3,3'-dipentyloxacarbocyanine iodide. Three ATCC reference strains of S. aureus and 13 unknown strains of the same microorganism were tested for susceptibility to penicillin G and oxacillin. Our results indicate that susceptibility of S. aureus to these antibiotics can be measured reliably at 90 min after addition of the antibiotic, and the results are comparable to those obtained with conventional susceptibility tests.

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“…[5][6][7] Given that a reduction of the time needed to perform an AST has been achieved, our method would improve patients' outcome, 2 could decrease the health care expenditure 18 and may delay the rise of bacterial resistances. 1 Wheat et al 7 tested 76 clinical isolates of Enterobacteriaceae to Ampicillin, Piperacillin and Gentamicin by ATP bioluminescence. They obtained satisfactory results for most organisms tested, except for P. mirabilis.…”

Section: Discussionmentioning

confidence: 99%

“…Moreover, non-fermenting gramnegative rods (NFGNR) still remain to be tested. [5][6][7] In this context, the aim of this work was to perform a rapid and complete AST using a firefly bioluminescent ATP assay in which the most predominant bacteria isolated in Clinical Microbiology laboratories were tested against some used antibiotics in clinical practice.…”

Section: Introductionmentioning

confidence: 99%