SummaryNinety-two consecutive patients referred for suspicion of deep venous thrombosis (DVT) were analyzed for D-dimer using ELISA, latex test, and a new immunofiltration method (NycoCard D-Dimer). Contrast venography verified the diagnosis in 40, and excluded the diagnosis in 52 patients. The sensitivity, negative predictive values, specificity and positive predictive values were, for ELISA 98%, 95%, 38% and 54, for NycoCard D-Dimer 100%, 100%, 42% and 57% and for the latex test 73%, 78%, 75%, and 69%, respectively. Sensitivity and specificity were inversely related with increasing pathological cutoff value. Comparison of test results by concentration category revealed a good agreement between ELISA and NycoCard D-Dimer, but to less extent between latex and the two other tests. It is concluded that NycoCard D-Dimer and D-dimer ELISA are well-suited as exclusion tests for DVT. A plasma sample is tested with NycoCard D-Dimer in less than 2 min. Thus, this test combines advantageous analytical properties comparable to the ELISA-test, with rapidity and simplicity comparable to the latex test.
The allergens of a crude D. farinae extract were identified by means of crossed radioimmunoelectrophoresis (CRIE). A total of 11 allergens were identified. They were characterized as three major, three intermediate and five minor allergens, based on radiostaining in CRIE. In addition to the radiostaining due to allergens, some misleading radiostaining was observed associated with some of the other antigens. A purified commercially available preparation, Spectralgen Mite D. farinae, was compared with the crude extract by means of immunoelectrophoretic techniques and radioallergosorbent test (RAST) based techniques. The purified preparation was found to be fully representative of the allergens in the crude extract. All 11 allergens were identified in the purified preparation. The purification ratio of the allergens varied from 2 to 7, while the total specific allergenic activity was approximately 6 times higher in the purified preparation than in the crude extract, as judged from RAST-inhibition studies.
In this immunofiltration assay of D-dimer in plasma samples, the antigens are captured by a monoclonal antibody on a porous membrane, and labeled with the same antibody conjugated to gold colloids. The assay time is < 2 min, and a color of intensity proportional to the concentration of D-dimer is left on the membrane. The reference range (mean +/- 2 SD) was 0.336 +/- 0.133 mg/L (n = 69). Linearity was found up to 10 mg/L. Comparison with ELISA results (x) for 198 patients' samples demonstrated a linear regression equation of y = 0.99(+/- 0.05)x + 0.68(+/- 0.07) and a mean square error of 0.503. Comparison of visual reading of the color signal (y) vs reflectometric measurements (x) for 220 patients' samples demonstrated a linear regression equation of y = 2.5(+/- 0.06)x -0.22(+/- 0.04) and a mean square error of 0.095. Bilirubin, hemoglobin, fibrinogen, soluble fibrin, and fibrinogen degradation products and freezing/thawing of samples did not interfere. Some interference from rheumatoid factor, heparin, and the presence of cells or large lipid particles was seen. The variance (CV) was 8-12% within run, 10-18% between runs, and 13-20% between persons. The new assay constitutes a rapid and reliable analytical tool combining simplicity equivalent to that of latex tests with analytical information approaching that of ELISA.
Twenty-five antigens were demonstrated in a crude Dermatophagoides farinae (DF) extract by means of crossed immunoelectrophoresis (CIE), using a rabbit anti-DF serum. With the same antiserum, four antigens were demonstrated in a scabies mite preparations, indicating an immunological relationship between these scabies antigens and some of the DF antigens. In tandem CIE experiments the DF antigens were found to be partly identical to three of the scabies antigens. No IgE activity against any of these scabies antigens was evident in pooled sera from patients with house dust mite allergy by crossed radioimmunoelectrophoresis. However, one such patient had IgE antibodies to a DF antigen and a partial identical scabies antigen.
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