A rapid semiautomated bioassay of gentamicin based on luciferase assay of bacterial adenosine triphosphate

Abstract

A rapid semiautomated bioassay of gentamicin in serum based on the effect of gentamicin on intracellular adenosine triphosphate (ATP) levels in bacterial cultures is presented. The ATP assay was performed with the firefly luciferin/luciferase system and results were compared with an agar diffusion technique. The lower limit of detection was set at 1 microgram/ml. The accuracy of the ATP assay expressed as a recovery of gentamicin over the range 1--8 microgram/ml was 98--103% and S.D. varied between 11-15%. The reproducibility was approximately within the same limits. Other antibiotics tested did not interfere with the determination of gentamicin. 112 clinical serum specimens containing gentamicin alone (N = 55) or in combination with other antibiotics (N = 57) were assayed and results with the two methods compared (r = 0.94). Volumes required were 0.1 ml serum and results were available within 3 h.