Performance of a fully automated fluorometric enzyme immunoassay for serum myoglobin measurement.

Abstract

An automated fluorometric enzyme immunoassay system for the determination of serum myoglobin has been recently developed. This method is based on the sandwich immunoassay and uses two mouse monoclonal antimyoglobin antibodies; the first one is complexed onto glass fiber paper and the second is conjugated to an enzyme alkaline phosphatase which reacts with the substrate 4-methylumbelliferyl phosphate to generate a fluorescent product. Using a dedicated automated apparatus the time to the first result is eight minutes, with additional values being produced at one-minute intervals (about 50 samples/hour). We compared the analytical performance of this fluorometric enzyme immunoassay with that of a RIA set up in our laboratory for the routine assay of serum myoglobin. The automated fluorometric enzyme immunoassay showed lower between-assay variability (CV = 4.7% vs 13.8%) and higher sensitivity (0.3 ng/mL vs 7.2 ng/mL) than the manual RIA. Moreover, the two immunoassays gave similar results when serum samples of normal subjects and patients with coronary artery disease with or without acute myocardial infarction (AMI) were assayed (fluorometric immunoassay = -0.7 + 0.851 RIA, r = 0.991, n = 137). In conclusion, the automated fluorometric enzyme immunoassay tested in the present study produces reliable clinical results with a rapid turnaround time and therefore can be recommended for use in the early detection of AMI in a laboratory of Coronary Care Unit.