Measurement of fibrin D-dimer may be a useful diagnostic test to exclude a diagnosis of deep venous thrombosis (DVT) in the emergency department setting. However, the specific assay format may influence its sensitivity and ultimate clinical utility. We tested samples from 200 patients under evaluation for DVT using three fibrin D-dimer assays: the SimpliRED whole blood agglutination assay, a latex agglutination assay, and the Dimertest EIA. Latex agglutination assays were performed in both a specialized laboratory and a routine laboratory. The negative predictive value for all tests was > 90%. The sensitivity of the SimpliRED assay was similar to that of the latex assay. The sensitivity of the latex assay was significantly lower when performed by generalist laboratory technologists. Thus, while D-dimer may be a useful test for the exclusion of DVT, subjective endpoint latex agglutination assays should be performed only by appropriately trained personnel.