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- Title
Novel method for simultaneously detecting HPA and HLA antibodies using Luminex microbeads.
- Authors
Tao, Sudan; Chen, Shu; Hong, Xiaozhen; He, Ji; Zhu, Faming
- Abstract
<bold>Background: </bold>Alloantibodies against human platelet antigens (HPAs) and human leukocyte antigen (HLA) are implicated in several immune-mediated platelet disorders. Detection of these antibodies is crucial in the diagnosis and management of these disorders. The aim of this study was to establish a novel method to simultaneously detect HPA-1, HPA-2, HPA-3, HPA-5 and HLA antibodies with Luminex microbeads technology.<bold>Methods: </bold>Monoclonal antibodies specific for platelet glycoproteins and HLA class I molecules were separately coupled to the Luminex microbeads. We validated specificity of the Luminex platform using the following antibodies: anti-HPA-1a, anti-HPA-2b, anti-HPA-3a, anti-HPA-5a, and anti-HLA positive samples. Sensitivity was evaluated by a serial dilution (from neat to 1/1024) using the following antibodies: anti-HPA-1a, anti-HPA-3a standard sera, and anti-HPA-5a positive serum. Serum samples were collected from 36 neonatal alloimmune thrombocytopenia (NAIT) patients suspected of having HPA or HLA antibodies and 8 samples from ISBT platelet workshop were tested using the Luminex assay.<bold>Results: </bold>The Luminex assay detected all antibodies tested from the known samples. The sensitivities of the Luminex assay detecting anti-HPA-1a, anti-HPA-3a, and anti-HPA-5a were 1:512, 1:64, and 1:128, respectively. The sensitivity of Luminex assay was higher than monoclonal antibody immobilization of platelet antigen method (MAIPA). No cross-reactivity was observed in the samples containing multi-platelet antibodies or mixture antibodies against HPA and HLA. The results of 44 samples with platelet disorders were consistent with those of the same samples processed with the MAIPA assay.<bold>Conclusion: </bold>Luminex microbeads coupled with monoclonal antibodies could be successfully used to detect HPA and HLA antibodies simultaneously, especially with high sensitivity in detecting HPA antibodies.
- Subjects
HISTOCOMPATIBILITY antigens; MONOCLONAL antibodies; IMMUNOGLOBULINS; BLOOD platelet disorders; BLOOD platelets; COMPARATIVE studies; IMMUNE system; IMMUNOASSAY; ISOANTIGENS; LATEX; RESEARCH methodology; MEDICAL cooperation; RESEARCH; THROMBOPENIC purpura; HLA-B27 antigen; EVALUATION research; GENOTYPES
- Publication
Journal of Translational Medicine, 2019, Vol 17, Issue 1, pN.PAG
- ISSN
1479-5876
- Publication type
journal article
- DOI
10.1186/s12967-019-2002-4