Abstract
Bacterial ATP levels in Escherichia coli cultures were determined by luciferin/luciferase assay. Rapid effects of gentamicin on ATP levels in the cultures were studied, and a dose-dependent accumulation of extracellular ATP was demonstrated. This phenomenon was used to develop a rapid bioassay of gentamicin in clinical serum specimens. The accuracy of the assay, expressed as coefficient of variation over the range 16 to 1 μg/ml, varied between 1.8 and 6.5%, compared to 2.3 to 4.7% for an agar disk diffusion assay. Serum gentamicin concentrations were determined by both methods in 122 serum specimens from 43 patients receiving gentamicin alone or in combination with other antibiotics, and the results were compared (r = 0.942). Determination of gentamicin based on luciferase assay of extracellular ATP in bacterial cultures requires only 10 μl of serum, allowing for capillary sampling, and results are available within 2 h.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Harber M. J., Asscher A. W. A new method for antibiotic assay based on measurement of bacterial adenosine triphosphate using the firefly bioluminescence system. J Antimicrob Chemother. 1977 Jan;3(1):35–41. doi: 10.1093/jac/3.1.35. [DOI] [PubMed] [Google Scholar]
- Lundin A., Richardsson A., Thore A. Continous monitoring of ATP-converting reactions by purified firefly luciferase. Anal Biochem. 1976 Oct;75(2):611–620. doi: 10.1016/0003-2697(76)90116-0. [DOI] [PubMed] [Google Scholar]
- Lundin A., Thore A. Analytical information obtainable by evaluation of the time course of firefly bioluminescence in the assay of ATP. Anal Biochem. 1975 May 26;66(1):47–63. doi: 10.1016/0003-2697(75)90723-x. [DOI] [PubMed] [Google Scholar]
- Lundin A., Thore A. Comparison of methods for extraction of bacterial adenine nucleotides determined by firefly assay. Appl Microbiol. 1975 Nov;30(5):713–721. doi: 10.1128/am.30.5.713-721.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nilsson L., Höjer H., Anséhn S., Thore A. A rapid semiautomated bioassay of gentamicin based on luciferase assay of bacterial adenosine triphosphate. Scand J Infect Dis. 1977;9(3):232–236. doi: 10.3109/inf.1977.9.issue-3.15. [DOI] [PubMed] [Google Scholar]
- Thore A., Nilsson L., Höjer H., Anséhn S., Bröte L. Effects of ampicillin on intracellular levels of adenosine triphosphate in bacterial cultures related to antibiotic susceptibility. Acta Pathol Microbiol Scand B. 1977 Apr;85(2):161–166. doi: 10.1111/j.1699-0463.1977.tb01690.x. [DOI] [PubMed] [Google Scholar]