Rapid determination of doxycycline based on luciferase assay of bacterial adenosine triphosphate

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Abstract
The effect of the bacteristatic antimicrobial doxycycline on intracellular adenosine triphosphate (ATP) levels in bacterial cultures was used to develop a rapid method to assay the drug in clinical serum specimens. ATP was quantified with the firefly luciferin/luciferase system. When compared with an agar disc diffusion technique results were well correlated within the range of 1 to 16 μg/ml. The accuracy of the luciferase assay expressed as recovery of doxycycline over the therapeutic range varied between 95 and 111% and a coefficient of variation of 4 to 15%. Seventy-six serum specimens from 46 patients were assayed and results with the two methods compared (r=0.89). Volumes required were 0.1 ml serum and results were available within 3 h.

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