Key result
The HPLC-ECD assay for platelet serotonin secretion was highly accurate compared to the reference isotopic test, with a bias of 1.48% and CI agreement of -18% to 15%.
Why the study?
Mild platelet secretion defects are challenging to diagnose, and most δ-granule secretion tests lack validation, are not quantitative, or have unreliable responses to weak agonists.
Does HPLC-ECD accurately measure platelet serotonin secretion compared to the reference isotopic test in subjects with platelet disorders?
Observational (n=87)
Does HPLC-ECD accurately measure platelet serotonin secretion compared to the reference isotopic test in subjects with platelet disorders?
HPLC-ECD is a highly accurate, quantitative, and sensitive alternative to the isotopic test for measuring platelet serotonin secretion in clinical laboratories.
Answers are grounded in this paper and cite their sources.
HPLC-ECD offers a validated non-isotopic option for platelet secretion testing; leaves open routine adoption pending larger clinical studies.
Background Mild secretion defects are the most frequent and challenging blood platelet disorders to diagnose. Most δ-granule secretion tests lack validation, are not quantitative, or have unreliable response to weak platelet agonists. Objectives To compare platelet serotonin secretion by HPLC-electrochemical detection technique (HPLC-ECD) with the reference isotopic test ( 3 H-5-HT), evaluating its performance in clinical laboratories. Methods The assay validation followed STARD-2015 recommendations. HPLC-ECD measured the nonsecreted serotonin remaining in platelet pellets after aggregation, comparing it with the reference 3 H-5-HT assay. We studied subjects with inherited and aspirin-induced blood platelet disorders and assessed the HPLC-ECD operation for routine clinical diagnosis. Results Calibration curves were linear (R 2 = 0.997), with SD for residuals of 3.91% and analytical sensitivity of 5ng/mL. Intra- and interassay imprecision bias ranged between −8.5% and 2.1% and −9% and 3.1%, respectively. Serotonin recovery and stability were >95%, and the variability range of measurements was −5.5% to 4.6%. Statistical differences detected between tests were biologically irrelevant, with bias of 1.48% (SD, 8.43) and CI agreement of −18% to 15%. Both assays distinctly detected platelet secretion induced by 10 μM epinephrine and 4 μmM adenosine diphosphate. However, HPLC-ECD is quantitative and more sensitive to low serotonin content in blood platelets. Reference cutoffs for each agonist were determined in 87 subjects. Initially, the HPLC-ECD requires relatively expensive equipment and trained operators but has remarkably cheap running costs and a turn-around time of 24-36 hours. We have used this diagnostic tool routinely for >8 years. Conclusion HPLC-ECD assay for platelet serotonin secretion is highly accurate, has advantages over the reference 3 H-5-HT test, and is suitable as a clinical laboratory technique.
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Aranda et al. (2023) conducted an observational in blood platelet disorders (n=87). HPLC-electrochemical detection technique (HPLC-ECD) vs. reference isotopic test (3H-5-HT) was evaluated on bias and CI agreement between tests (bias 1.48%, 95% CI -18 to 15). The HPLC-ECD assay for platelet serotonin secretion was highly accurate compared to the reference isotopic test, with a bias of 1.48% and CI agreement of -18% to 15%.
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