A rapid and non-invasive proteomic analysis using DBS and buccal swab for multiplexed second-tier screening of Pompe disease and Mucopolysaccharidosis type I.

Autor: Zhang T; Seattle Children's Research Institute, Seattle, WA, United States of America., Duong P; Seattle Children's Research Institute, Seattle, WA, United States of America., Dayuha R; Seattle Children's Research Institute, Seattle, WA, United States of America., Collins CJ; Seattle Children's Research Institute, Seattle, WA, United States of America., Beckman E; Division of Genetic Medicine, Seattle Children's Hospital, Seattle, WA, United States of America., Thies J; Division of Genetic Medicine, Seattle Children's Hospital, Seattle, WA, United States of America., Chang I; Biochemical Genetics Clinic, Seattle Children's Hospital, Seattle, WA, United States of America; Department of Pediatrics, Division of Genetic Medicine, University of Washington School of Medicine, Seattle, WA, United States of America., Lam C; Biochemical Genetics Clinic, Seattle Children's Hospital, Seattle, WA, United States of America; Department of Pediatrics, Division of Genetic Medicine, University of Washington School of Medicine, Seattle, WA, United States of America., Sun A; Biochemical Genetics Clinic, Seattle Children's Hospital, Seattle, WA, United States of America; Department of Pediatrics, Division of Genetic Medicine, University of Washington School of Medicine, Seattle, WA, United States of America., Scott AI; Department of Laboratory, Seattle Children's Hospital, Seattle, WA, United States of America., Thompson J; WA State Department of Health, Seattle, WA, United States of America., Singh A; WA State Department of Health, Seattle, WA, United States of America., Khaledi H; Department of Chemistry, University of Washington, Seattle, WA, United States of America., Gelb MH; Department of Chemistry, University of Washington, Seattle, WA, United States of America., Hahn SH; Seattle Children's Research Institute, Seattle, WA, United States of America; Biochemical Genetics Clinic, Seattle Children's Hospital, Seattle, WA, United States of America; Department of Pediatrics, Division of Genetic Medicine, University of Washington School of Medicine, Seattle, WA, United States of America. Electronic address: sihoun.hahn@seattlechildrens.org.
Jazyk: angličtina
Zdroj: Molecular genetics and metabolism [Mol Genet Metab] 2022 Aug; Vol. 136 (4), pp. 296-305. Date of Electronic Publication: 2022 Jun 28.
DOI: 10.1016/j.ymgme.2022.06.006
Abstrakt: Purpose: Current newborn screening programs for Pompe disease (PD) and mucopolysaccharidosis type I (MPS I) suffer from a high false positive rate and long turnaround time for clinical follow up. This study aimed to develop a novel proteomics-based assay for rapid and accurate second-tier screening of PD and MPS I. A fast turnaround assay would enable the identification of severe cases who need immediate clinical follow up and treatment.
Methods: We developed an immunocapture coupled with mass spectrometry-based proteomics (Immuno-SRM) assay to quantify GAA and IDUA proteins in dried blood spots (DBS) and buccal swabs. Sensitivity, linearity, reproducibility, and protein concentration range in healthy control samples were determined. Clinical performance was evaluated in known PD and MPS I patients as well as pseudodeficiency and carrier cases.
Results: Using three 3.2 mm punches (~13.1 μL of blood) of DBS, the assay showed reproducible and sensitive quantification of GAA and IDUA. Both proteins can also be quantified in buccal swabs with high reproducibility and sensitivity. Infantile onset Pompe disease (IOPD) and severe MPS I cases are readily identifiable due to the absence of GAA and IDUA, respectively. In addition, late onset Pompe disease (LOPD) and attenuated MPS I patients showed much reduced levels of the target protein. By contrast, pseudodeficiency and carrier cases exhibited significant higher target protein levels compared to true patients.
Conclusion: Direct quantification of endogenous GAA and IDUA peptides in DBS by Immuno-SRM can be used for second-tier screening to rapidly identify severe PD and MPS I patients with a turnaround time of <1 week. Such patients could benefit from immediate clinical follow up and possibly earlier treatment.
(Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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