In:
Clinical Chemistry and Laboratory Medicine (CCLM), Walter de Gruyter GmbH, Vol. 61, No. 6 ( 2023-05-25), p. 1075-1083
Abstract:
Traditional methods for β-thalassemia screening usually rely on the structural integrity of hemoglobin (Hb), which can be affected by the hemolysis of red blood cells and Hb degradation. Here, we aim to develop a reliable and high throughput method for rapid detection of β-thalassemia using dried blood spots (DBS). Methods Hb components were extracted from a disc (3.2 mm diameter) punched from the DBS samples and digested by trypsin to produce a series of Hb-specific peptides. An analytical system combining high-resolution mass spectrometry and high-performance liquid chromatography was used for biomarker selection. The selected marker peptides were used to calculate delta/beta (δ/β) and beta-mutated/beta (β M /β) globin ratios for disease evaluation. Results Totally, 699 patients and 629 normal individuals, aged 3 days to 89 years, were recruited for method construction. Method assessment showed both the inter-assay and intra-assay relative standard deviation values were less than 10.8%, and the limits of quantitation for the proteo-specific peptides were quite low (1.0–5.0 μg/L). No appreciable matrix effects or carryover rates were observed. The extraction recoveries ranged from 93.8 to 128.7%, and the method was shown to be stable even when the samples were stored for 24 days. Prospective applications of this method in 909 participants also indicated good performance with a sensitivity of 100% and a specificity of 99.6%. Conclusions We have developed a fast, high throughput and reliable method for screening of β-thalassemia and hemoglobinopathy in children and adults, which is expected to be used as a first-line screening assay.
Type of Medium:
Online Resource
ISSN:
1434-6621
,
1437-4331
DOI:
10.1515/cclm-2022-0706
Language:
English
Publisher:
Walter de Gruyter GmbH
Publication Date:
2023
detail.hit.zdb_id:
1492732-9
SSG:
15,3
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