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Published on: August 8, 2013
Chemiluminescence resonance energy transfer-based method to investigate the platelet surface molecule in acute
Linyan He1, Bing Ding2, Fei Shen1
1Jiangsu Institute of Hematology, NHC Key Laboratory of Thrombosis and Hemostasis, National Clinical Research Center for Hematologic Diseases, Collaborative Innovation Center of Hematology, Suzhou, Jiangsu, PR China.
Insights
A new chemiluminescence resonance energy transfer (CRET) assay effectively detects platelet activation markers CD61 and CD62P. This method shows higher CD62P levels in acute myocardial infarction (AMI) patients, offering a novel approach for diagnosing AMI.
Area of Science:
- Biochemistry and Molecular Biology
- Cardiovascular Medicine
- Biomedical Diagnostics
Background:
- Platelet activation plays a crucial role in acute myocardial infarction (AMI) pathogenesis.
- Accurate and rapid detection of platelet activation markers is essential for AMI diagnosis and management.
- Existing methods for detecting platelet surface markers may have limitations in speed or sensitivity.
Purpose of the Study:
- To evaluate the efficacy of a novel chemiluminescence resonance energy transfer (CRET)-based assay for detecting platelet activation markers CD61 and CD62P (P-selectin).
- To compare platelet marker expression in patients with AMI versus healthy controls.
- To assess the correlation between CRET assay results and traditional methods like flow cytometry.
Main Methods:
- A CRET-based assay was developed and validated for quantifying CD61 and CD62P expression on platelets.
- Blood samples were collected from 22 AMI patients and 22 matched healthy controls.
- Platelet activation was induced in vitro using agonists like arachidonic acid (AA) and adenosine diphosphate (ADP).
Main Results:
- The CRET assay demonstrated high precision (low coefficient of variation) and excellent linearity (R² > 0.995) for CD61 and CD62P detection.
- AMI patients exhibited significantly higher basal CD62P levels compared to healthy controls, with a strong positive correlation between platelet aggregation rate and CD62P expression (r=0.8354, p<.0001).
- CRET assay results for CD61 and CD62P showed good correlation with flow cytometry measurements, both at baseline and after agonist stimulation.
Conclusions:
- The CRET-based assay is a reliable and sensitive method for detecting platelet activation markers CD61 and CD62P.
- Elevated CD62P expression is a significant indicator of platelet activation in AMI patients.
- This one-step CRET assay offers a promising new diagnostic tool for assessing platelet reactivity in acute myocardial infarction.
Abstract:
The chemiluminescence resonance energy transfer (CRET)-based method was applied to the detect of CD61 and CD62P (P-selectin) on the surface of platelet in patients with acute myocardial infarction (AMI) and healthy controls. Patients admitted to the Department of Emergency in The First Affiliated Hospital of Soochow University from May 2025 to December 2025 were recruited. We studied 22 AMI patients who were matched with 22 controls. The coefficient of variations and standard curves of indexes were calculated. Expressions of CD61 and CD62P in plasma were evaluated by CRET assay. Standard curve property for CD61 and CD62P: R2 .995 and 0.9999. The coefficient of variation in CRET system was 3.39%, 1.76% (QC high), 6.25% and 7.16% (QC low) for CD61 and CD62P, respectively. CRET data showed the agonist (arachidonic acid and ADP) concentration dependency on platelet activation. AMI patients had higher basal CD62P levels when compared to controls. The platelet aggregation rate of AMI patients had a strong positive correlation with the expression of CD62P (r = 0.8354, p < .0001). In the healthy controls, the platelet CD61 and CD62P expression detected by flow cytometry showed a positive correlation with that evaluated by CRET (r = 0.4296, p = .0460; r = 0.5662, p = .0060, respectively). After platelet activation with AA or ADP, the expression levels of CD62P detected by flow cytometry were positively correlated with those measured by the CRET method (r = 0.6195, p = .0021; r = 0.7663, p < .0001, respectively). Expressions of CD62P elevated after agonist stimulation in AMI patients and controls in vitro. With the biomarker of CD62P as the model, the Area Under Curve (AUC) was 0.9236 (p < .0001). The cutoff value of CD62P to evaluate platelet reactivity by CRET was 3.985%. This one-step CERT assay provided a new approach to detect platelet activity of acute myocardial infarction.
