Unexpected suppression of total digoxin concentrations by cross-reactants in the microparticle enzyme immunoassay: elimination of interference by monitoring free digoxin concentration.
The microparticle enzyme immunoassay (MEIA for digoxin (Abbott Laboratories, Abbott Park, Ill) requires no sample pretreatment and is widely used in clinical toxicology laboratories for monitoring serum digoxin concentrations. One advantage of the new MEIA is the lower cross-reactivities with such cross-reactants as digitoxin, oleandrin, and bufalin compared with the fluorescence polarization immunoassay (FPIA)for digoxin. Digitoxin, oleandrin, and bufalin showed positive cross-reactivity with MEIA and FPIAs for digoxin in the absence of the primary analyte, digoxin. A surprising finding was that digoxin concentrations were falsely decreased by these cross-reactants when serum pools containing digoxin were supplemented with various concentrations of these cross-reactants and when digoxin concentrations were measured by the MEIA. In contrast, digoxin concentrations were falsely elevated when measured by the FPIA. For example, when a serum pool containing 2.15 nmol/L of digoxin was supplemented with 129.5 nmol/L of bufalin, the apparent digoxin concentrations were 1.45 nmol/L with the MEIA and 3.00 nmol/L with the FPIA. Taking the advantage of only 25% protein binding of digoxin and more than 95% protein binding of digitoxin and bufalin, we demonstrated that monitoring free digoxin instead of total digoxin eliminated negative interference of digoxin by these cross-reactants in the MEIA and positive interference in the FPIA. Although oleandrin is also strongly bound to serum protein, high concentrations of oleandrin still modestly affect the free digoxin assay for both MEIA and FPIAs.
Dasgupta
A
A
Department of Pathology, University of New Mexico Health Sciences Center, Albuquerque, USA.
Scott
J
J
D002018:Bufanolides / Q000097:blood; D002298:Cardenolides / Q000097:blood; D002301:Cardiac Glycosides / Q000276:immunology; D002316:Cardiotonic Agents / Q000097:blood*; D003429:Cross Reactions; D004074:Digitoxin / Q000097:blood; D004077:Digoxin / Q000097:blood*; D005189:False Positive Reactions; D016231:Fluorescence Polarization Immunoassay; D006801:Humans; D007124:Immunoenzyme Techniques; D008863:Microspheres
D002018:Bufanolides; D002298:Cardenolides; D002301:Cardiac Glycosides; D002316:Cardiotonic Agents; D004077:Digoxin; D004074:Digitoxin; C021065:oleandrin; C022777:bufalin