An improved application for the enzyme multipled immunoassay technique for caffeine, amikacin, and methotrexate assays on the Dade-Behring dimension RxL Max clinical chemistry system

Caffeine is widely used in children's hospitals to treat neonatal apnea. Amikacin is used for treating hospital-acquired infections caused by Gram-negative bacteria resistant to other aminoglycosides. The blood levels, however, have to be monitored carefully because of its ototoxicity and nephrotoxicity. Methotrexate (MTX) is used as a chemotherapeutic agent in the treatment of leukemia and lymphoma as well as of certain solid tumors. Current enzyme multiplied immunoassay technique (EMIT) assays for caffeine, amikacin, and MTX lack low-end precision. In addition, the EMIT assays for MTX lack the sensitivity of reliable quantification to 0.05 μmol/L needed because of today's more rigorous requirements. The goal of the present study was to optimize the EMIT method parameters on the Dimension RxL Max, thereby providing applications with improved precision for all the three analytes and enhancing the sensitivity of the EMIT methotrexate assay. Serum samples were measured for caffeine, amikacin, and MTX by EMIT on the Dimension RxL Max and by EMIT (on the Olympus AU 600 for caffeine) and fluorescence polarization immunoassay [for MTX and amikacin (FPIA; TDx FLx)] at Quest Diagnostics. The new instrument method parameters that use larger sample volumes and longer observation of optical density changes (caffeine, MTX) provide improved sensitivity for MTX permitting reliable measurement at 0.05 μmol/L and improved precision for all three analytes. Within- and between-day imprecision were less than 6% for low to high concentrations of caffeine and amikacin controls and are less than 7.5% for MTX concentrations greater than 0.3 μmol/L and 12.3% at 0.06 μmol/L. The correlation coefficients for caffeine, amikacin, and MTX plotted for the Dimension RxL Max versus the methods used at Quest Diagnostics were 0.973, 0.986, and 0.992, respectively. These EMIT method applications now compare well with other established assays. The new Dimension RxL Max method parameters provide greatly improved precision and also meet today's clinical sensitivity guidelines (0.05 μmol/L) for MTX.