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Enzyme activity in dried blood spot as a diagnostic tool for adenosine deaminase 2 deficiency
- Source :
- Analytical Biochemistry. 628:114292
- Publication Year :
- 2021
- Publisher :
- Elsevier BV, 2021.
-
Abstract
- Background Deficiency of adenosine deaminase 2 (DADA2) is an autoinflammatory disease caused by mutations in the adenosine deaminase 2 (ADA2) gene. Loss of functional ADA2 activity results in vasculitis syndrome, immunodeficiency, and hematopoietic disorders. Early diagnosis is required for effective treatment. Methods We developed a dried blood spot (DBS)-based ADA2 activity colorimetric assay. Heparin-affinity purification was used during sample preparation to improve the assay more efficiently. The stability of ADA2 during DBS storage and ADA2 activity of DADA2 patients and healthy controls were examined. Results Active ADA2 was extracted from the DBS of healthy controls. ADA2 activity in DBS, stored either frozen or refrigerated, remained stable for at least 90 days. A significant difference in ADA2 activity was observed between healthy controls and patients. No ADA2 activity was detected in DBS from patients. Conclusions Our new DBS ADA2 activity assay is experimentally simple, highly adaptable, and requires no special equipment except for a microplate reader. A low background was achieved with heparin-affinity purification. The method differentiates clearly between healthy controls and patients. ADA2 activity can be reliably measured in DBS, providing an opportunity to diagnose DADA2 at an early stage.
- Subjects :
- Adult
Adenosine Deaminase 2 Deficiency
Adolescent
Adenosine Deaminase
Biophysics
Pharmacology
01 natural sciences
Biochemistry
Young Adult
03 medical and health sciences
medicine
Humans
Child
Molecular Biology
Immunodeficiency
030304 developmental biology
0303 health sciences
biology
business.industry
010401 analytical chemistry
Infant, Newborn
Infant
Cell Biology
Heparin
ADENOSINE DEAMINASE 2
medicine.disease
Enzyme assay
0104 chemical sciences
Dried blood spot
Haematopoiesis
biology.protein
Intercellular Signaling Peptides and Proteins
Dried Blood Spot Testing
Vasculitis
business
medicine.drug
Subjects
Details
- ISSN :
- 00032697
- Volume :
- 628
- Database :
- OpenAIRE
- Journal :
- Analytical Biochemistry
- Accession number :
- edsair.doi.dedup.....b4ac9038525f5c3da3b8b472cd57b0f9