Korean J Lab Med.  2006 Jun;26(3):192-197. 10.3343/kjlm.2006.26.3.192.

Detection of Platelet Specific Antibodies by Modified Antigen Capture ELISA Test

• Kim H
• Oh EJ
• Kim J
• Park YJ
• Han K

• KMID: 2238886
• DOI: http://doi.org/10.3343/kjlm.2006.26.3.192

Abstract

BACKGROUND: Autoimmune thrombocytopenia (AITP) is characterized by autoantibody-induced platelet destruction. Although several studies have shown that pathogenic autoantibodies are mainly IgG directed platelet glycoproteins (GP), a platelet GP specific test is not available in clinical laboratories. The aim of this study was to evaluate the clinical usefulness of a Modified Antigen Capture Enzyme-linked immunosorbent assay (MACE) test in the diagnosis of AITP.
METHODS
We investigated fifty-seven patients who showed a platelet count lower than 100 x 10(9)/L and underwent a bone marrow examination. They were classified into primary AITP (P-AITP) (n=21), secondary AITP (S-AITP) (n=15), and non-immune thrombocytopenia (NITP) (n=21) by bone marrow findings and clinical diagnosis. Platelet GP (IIb/IIIa, Ia/IIa, Ib/IX, IV)-specific antibodies and anti-HLA class I antibody were detected by MACE test.
RESULTS
Among 57 samples, platelet GP specific antibodies were detected in 8 (22.2%) of 36 patients with AITP and 1 (4.8%) of 21 patients with NITP. The specificities were as follows: GP IIb/IIIa (n=4), GP Ia/IIa (n=5), GP Ib/IX (n=3) and GPIV (n=2). Of the nine patients with platelet GP specific antibodies, four (44.4%) had more than two platelet GP specific antibodies. The sensitivity, specificity, positive predictive value and negative predictive values of the MACE test for AITP were 22.2%, 95.2%, 88.9%, 41.7%, respectively. A previous transfusion history was associated with a higher detection rate of anti-HLA class I antibodies (P<0.05).
CONCLUSIONS
The MACE test is a convenient method to detect platelet GP specific antibody and is very specific to diagnose AITP. In clinical practice, even though it is not sensitive, the MACE test would be useful in differentiating AITP from NITP.

Keyword

Autoimmune thrombocytopenia; Modified Antigen Capture ELISA test (MACE); Platelet glycoprotein specific antibody; Antiplatelet antibodies

MeSH Terms

Antibodies*
Autoantibodies
Blood Platelets*
Bone Marrow
Bone Marrow Examination
Diagnosis
Enzyme-Linked Immunosorbent Assay*
Humans
Immunoglobulin G
Platelet Count
Platelet Membrane Glycoproteins
Purpura, Thrombocytopenic, Idiopathic
Sensitivity and Specificity
Thrombocytopenia
Antibodies
Autoantibodies
Immunoglobulin G
Platelet Membrane Glycoproteins

Figure

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  Fig. 1. Detection of platelet specific IgG antibodies to platelet glycoproteins (GP) IIb/IIIa, Ia/IIa, Ib/IX, IV or HLA class I (GP IIb/IIIa in this example) by MACE assay. Washed platelets were first incubated with human plasma (containing anti-platelet GP IIb/IIIa in this sample). The antibody-sensitized platelets were then lysed and the supernatant was added to a microplate precoated with murine monoclonal antibodies (MoAb) specific to platelet GP or HLA class I. Any human IgG bound to the GP or HLA class I antigen were detected with a alkaline phosphatase (ALP) conjugated antihuman IgG and substrate solution.

Reference

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