• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of aacPermissionsJournals.ASM.orgJournalAAC ArticleJournal InfoAuthorsReviewers
Antimicrob Agents Chemother. Aug 1980; 18(2): 264–268.
PMCID: PMC283981

Substrate-labeled fluorescent immunoassay for amikacin in human serum.

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.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (708K), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Barza M, Scheife RT. 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]
  • Burd JF, Wong RC, Feeney JE, Carrico RJ, Boguslaski RC. Homogeneous reactant-labeled fluorescent immunoassay for therapeutic drugs exemplified by gentamicin determination in human serum. Clin Chem. 1977 Aug;23(8):1402–1408. [PubMed]
  • Lerner SA, Seligsohn R, Matz GJ. Comparative clinical studies of ototoxicity and nephrotoxicity of amikacin and gentamicin. Am J Med. 1977 Jun;62(6):919–923. [PubMed]
  • Lewis JE, Nelson JC, Elder HA. Radioimmunoassay of an antibiotic: gentamicin. Nat New Biol. 1972 Oct 18;239(94):214–216. [PubMed]
  • Lewis JE, Nelson JC, Elder HA. Amikacin: a rapid and sensitive radioimmunoassay. Antimicrob Agents Chemother. 1975 Jan;7(1):42–45. [PMC free article] [PubMed]
  • Maitra SK, Yoshikawa TT, Steyn CM, Guze LB, Schotz MC. Amikacin assay in serum by high-performance liquid chromatography. Antimicrob Agents Chemother. 1978 Dec;14(6):880–885. [PMC free article] [PubMed]
  • Marengo PB, Wilkins J, Overturf GD. Rapid, specific microbiological assay for amikacin (BB-K8). Antimicrob Agents Chemother. 1974 Oct;6(4):498–500. [PMC free article] [PubMed]
  • Scarbrough E, Williams JW, Northrop DB. Spectrophotometric assay for amikacin using purified kanamycin acetyltransferase. Antimicrob Agents Chemother. 1979 Aug;16(2):221–224. [PMC free article] [PubMed]
  • Stevens P, Young LS, Hewitt WL. 125I-Radioimmunoassay of amikacin and comparison with a microbioassay. J Antibiot (Tokyo) 1976 Aug;29(8):829–832. [PubMed]
  • Wong RC, Burd JF, Carrico RJ, Buckler RT, Thoma J, Boguslaski RC. Substrate-labeled fluorescent immunoassay for phenytoin in human serum. Clin Chem. 1979 May;25(5):686–691. [PubMed]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...