Abstract
A homogeneous substrate-labeled fluorescent immunoassay has been developed to measure amikacin levels in human serum. Amikacin is covalently labeled with the fluorogenic enzyme substrate beta-galactosyl-umbelliferone. This beta-galactosyl-umbelliferone-amikacin conjugate is nonfluorescent under assay conditions until it is hydrolyzed by beta-galactosidase to yield a fluorescent product. When antiserum to amikacin binds the substrate-labeled drug, the antibody complex formation inhibits hydrolysis of the fluorogenic substrate. Reaction mixtures containing a constant level of substrate-labeled amikacin and a limiting amount of antiserum enable labeled and unlabeled amikacin to compete for the antibody-binding sites. Unbound substrate-labeled drug is hydrolyzed by the enzyme to release a fluorescent product that is proportional to the unlabeled amikacin concentration. The amikacin levels found in clinical serum samples with this method were comparable (r = 0.987) to those obtained by radioimmunoassay. The fluorescent immunoassay is rapid and simple to perform and requires only 2 microliters of serum.
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Selected References
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- Barza M., Scheife R. T. Drug therapy reviews: Antimicrobial spectrum, pharmacology and therapeutic use of antibiotics--part 4: aminoglycosides. Am J Hosp Pharm. 1977 Jul;34(7):723–737. [PubMed] [Google Scholar]
- Burd J. F., Wong R. C., Feeney J. E., Carrico R. J., Boguslaski R. C. Homogeneous reactant-labeled fluorescent immunoassay for therapeutic drugs exemplified by gentamicin determination in human serum. Clin Chem. 1977 Aug;23(8):1402–1408. [PubMed] [Google Scholar]
- Lerner S. A., Seligsohn R., Matz G. J. Comparative clinical studies of ototoxicity and nephrotoxicity of amikacin and gentamicin. Am J Med. 1977 Jun;62(6):919–923. doi: 10.1016/0002-9343(77)90661-1. [DOI] [PubMed] [Google Scholar]
- Lewis J. E., Nelson J. C., Elder H. A. Amikacin: a rapid and sensitive radioimmunoassay. Antimicrob Agents Chemother. 1975 Jan;7(1):42–45. doi: 10.1128/aac.7.1.42. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lewis J. E., Nelson J. C., Elder H. A. Radioimmunoassay of an antibiotic: gentamicin. Nat New Biol. 1972 Oct 18;239(94):214–216. doi: 10.1038/newbio239214a0. [DOI] [PubMed] [Google Scholar]
- Maitra S. K., Yoshikawa T. T., Steyn C. M., Guze L. B., Schotz M. C. Amikacin assay in serum by high-performance liquid chromatography. Antimicrob Agents Chemother. 1978 Dec;14(6):880–885. doi: 10.1128/aac.14.6.880. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marengo P. B., Wilkins J., Overturf G. D. Rapid, specific microbiological assay for amikacin (BB-K8). Antimicrob Agents Chemother. 1974 Oct;6(4):498–500. doi: 10.1128/aac.6.4.498. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scarbrough E., Williams J. W., Northrop D. B. Spectrophotometric assay for amikacin using purified kanamycin acetyltransferase. Antimicrob Agents Chemother. 1979 Aug;16(2):221–224. doi: 10.1128/aac.16.2.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stevens P., Young L. S., Hewitt W. L. 125I-Radioimmunoassay of amikacin and comparison with a microbioassay. J Antibiot (Tokyo) 1976 Aug;29(8):829–832. doi: 10.7164/antibiotics.29.829. [DOI] [PubMed] [Google Scholar]
- Wong R. C., Burd J. F., Carrico R. J., Buckler R. T., Thoma J., Boguslaski R. C. Substrate-labeled fluorescent immunoassay for phenytoin in human serum. Clin Chem. 1979 May;25(5):686–691. [PubMed] [Google Scholar]