Cumulative experience with a simplified solid-phase radioimmunoassay for the detection of bound antiplatelet IgG, serum auto-, allo-, and drug-dependent antibodies
A simplified, sensitive, solid-phase radioimmunoassay employing /sup 125/I-staphylococcal protein A has been developed that is capable of detecting bound antiplatelet IgG as well as serum auto-, allo-, and drug-dependent antiplatelet antibodies. The simplified assay employs a ratio of test over control platelet counts per minute (cpm) for detection of positive results. All reagents are commercially available. The assay can be performed with as little as 10/sup 6/ washed platelets (10 ..mu..l of whole blood) that have been stored for as long as 8 wk at 4/sup 0/C in microtiter plates. The assay time, employing stored platelets, is 4 hr. Bound platelet IgG is positive in 93% of 46 thrombocytopenic patients with autoimmune disease and correlates inversely with their platelet count, r = -0.65, p < 0.001. The ability of this assay to detect serum antibody was studied with a rabbit anti-human platelet antibody capable of giving optimal immunoprecipitation with solubilized platelet membranes at a titer of 1:10. The present assay increases the sensitivity of antibody detection 256-fold to a titer of 1:2560. Human serum antiplatelet membrane antibody was positive in 2 of 2 patients with anti-PLA-1 antibody (titers of 1:256 and >1:64); 7 of 12 multiply transfused patients who were refractory to platelet transfusion (2 had titers of >1:256 and 1:32); 5 of 19 patients with autoimmune thrombocytopenic purpura (2 had titers of 1:64 and 1:32); and 10 of 14 patients with clinical histories of drug-dependent antiplatelet antibody (2 had titers of 1:1280 for quinidine and 1:384 for phenazopyridine).
- Research Organization:
- New York Univ. Medical Center, NY
- OSTI ID:
- 6172850
- Journal Information:
- Blood; (United States), Vol. 60:4
- Country of Publication:
- United States
- Language:
- English
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550601* - Medicine- Unsealed Radionuclides in Diagnostics