Back to Search
Start Over
High-Throughput CD36 Phenotyping on Human Platelets Based on Sandwich ELISA and Mutant Gene Analysis.
- Source :
-
Transfusion Medicine & Hemotherapy . 2024, Vol. 51 Issue 1, p32-40. 9p. - Publication Year :
- 2024
-
Abstract
- Background: CD36 deficiency is closely associated with fetal/neonatal alloimmune thrombocytopenia, platelet transfusion refractoriness, and other hemorrhage disorders, particularly in Asian and African populations. There is a clinical need for rapid and high-throughput methods of platelet CD36 (pCD36) phenotyping to improve the availability of CD36 typing of donors and assist clinical blood transfusions for patients with anti-CD36 antibodies. Such methods can also support the establishment of databases of pCD36-negative phenotypes. Study Design and Methods: A sandwich enzyme-linked immunosorbent assay (ELISA) for CD36 phenotyping of human platelets was developed using anti-CD36 monoclonal antibodies. The reliability of the assay was evaluated by calculating the intra-assay and inter-assay coefficients of variation (CV). A total of 1,691 anticoagulant whole blood samples from healthy blood donors were randomly selected. PCD36 expression was measured using a sandwich ELISA. PCD36 deficiency was confirmed by flow cytometry (FC). Mutations underlying pCD36 deficiency were identified using polymerase chain reaction sequence-based typing (PCR-SBT). Results: The sandwich ELISA for pCD36 phenotyping had high reliability (intra-assay CV, 2.1–4.8%; inter-assay CV, 2.3–5.2%). The sandwich ELISA was used to screen for CD36 expression on platelets isolated from 1,691 healthy blood donors. Of these, 36 samples were pCD36-negative. FC demonstrated absence of CD36 expression on monocytes in three of the 36 cases. In the present study population, the frequency of CD36 deficiency was 2.13% (36/1,691), of which 0.18% (3/1,691) was type I deficiency and 1.95% (33/1,691) was type II deficiency. In addition, we used PCR-SBT to characterize the gene mutations in exons 3–14 of the CD36 gene in 27 cases of CD36 deficiency and discovered 10 types of mutations in 13 pCD36-negative samples. Conclusion: The present study describes the development and characterization of a highly reliable sandwich ELISA for high-throughput screening for pCD36 expression. This novel method is feasible for clinical applications and provides a useful tool for the establishment of databases of pCD36-negative phenotype donors. [ABSTRACT FROM AUTHOR]
- Subjects :
- *ANTIGEN analysis
*HIGH throughput screening (Drug development)
*FLOW cytometry
*IMMUNOGLOBULINS
*GENETIC mutation
*BLOOD platelets
*BLOOD transfusion
*GENE expression
*COMPARATIVE studies
*ENZYME-linked immunosorbent assay
*RESEARCH funding
*DESCRIPTIVE statistics
*POLYMERASE chain reaction
*STATISTICAL sampling
*DATA analysis software
*GENETIC techniques
*PHENOTYPES
Subjects
Details
- Language :
- English
- ISSN :
- 16603796
- Volume :
- 51
- Issue :
- 1
- Database :
- Academic Search Index
- Journal :
- Transfusion Medicine & Hemotherapy
- Publication Type :
- Academic Journal
- Accession number :
- 175341821
- Full Text :
- https://doi.org/10.1159/000530039