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Antimicrob Agents Chemother. 1975 January; 7(1): 42-45
Copyright © 1975 American Society for Microbiology. All Rights Reserved.

Amikacin: a Rapid and Sensitive Radioimmunoassay

John E. Lewis, Jerald C. Nelson and Harvey A. Elder

Departments of Medicine and Radiology, Loma Linda University Medical Center, Loma Linda, California 92354

ABSTRACT

A rapid, sensitive, and specific radioimmunoassay has been developed for the measurement of amikacin, a new semisynthetic aminoglycoside antibiotic. Antisera were produced by immunizing rabbits with amikacin conjugated to porcine thyroglobulin. Sera were screened for amikacin antibodies with 3H-labeled amikacin. Free and bound radioactivity were separated by the second antibody method. As little as 5 ng of the antibiotic can be measured in serum, cerebrospinal fluid, and urine. The standard curve is unaffected by pH (5–11), ionic strength (0.01 M to 0.3 M), MgCl2 or CaCl2 (1 to 8 mM/liter), or urea (10–300 mg/100 ml). The antibodies raised to amikacin are not able to distinguish between kanamycin and amikacin. The cross reactivity of all other antibiotics tested was <0.1%. Co-incubation of first and second antibody allows for a 30-min incubation period, making the assay useful for stat determinations. The standard curve is linear on a logit-log plot, allowing for rapid computer analysis of the standard curve, interpolation of sample values, and quality-control monitoring.

Antimicrob Agents Chemother. 1975 January; 7(1): 42-45
Copyright © 1975 American Society for Microbiology. All Rights Reserved.





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